Embed
Email

MS Excel file

Document Sample

Shared by: linzhengnd
Categories
Tags
Stats
views:
0
posted:
11/8/2011
language:
English
pages:
58
Page 1 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

Accession # 1 Reference

NGV4(from NGC) Lin et al. 1994

DQ518643

P-GV3(from PUO-218) Lin et al. 1994

DQ518667

AY648301 rDEN4 Blaney et al. 2003

AY648301 rDEN4 Blaney et al. 2003

AY648301 rDEN4 Blaney et al. 2003

AY648301 rDEN4 Blaney et al. 2003

AY648301 rDEN4 Blaney et al. 2003

AY648301 rDEN4 Blaney et al. 2003

AY648301 rDEN4 Blaney et al. 2003

AY648301 rDEN4 Blaney et al. 2003

AY648301 rDEN4 Blaney et al. 2003

AY648301 rDEN4 Blaney et al. 2003

TBE V-IC3 Holzmann et al. 1997

U39292

U39292 TBE V-IE3 Holzmann et al. 1997

U39292 TBE V-IO3 Holzmann et al. 1997

not identified WNI-568 Chambers et al. 1998



not identified WNI-567 Chambers et al. 1998

not identified WNI-25 Chambers et al. 1998

not identified WNI-25A Chambers et al. 1998

not identified DEN3 via Mab 1H9 Serafin & Aaskov 2001



PRS 225489 DEN2 (NGC) Via Mab 4G2 Serafin & Aaskov 2001



M29095 DEN2 Mexican Small plaque 3 Sanchez & Ruiz 1996



L04561 DEN2 Mexican Medium plaque 1 Sanchez & Ruiz 1996



L04561 DEN2 Mexican Large plaque 2 Sanchez & Ruiz 1996

L04561 DEN2 Mexican Small plaque 2 Sanchez & Ruiz 1996



L04561 DEN2 Mexican Small plaque 1 Sanchez & Ruiz 1996



L04561 DEN2 Mexican Small plaque 4 Sanchez & Ruiz 1996



L04561 DEN2 Mexican Large plaque 3 Sanchez & Ruiz 1996

L04561 DEN2 Mexican Large plaque 4 Sanchez & Ruiz 1996

not identified DEN Roehrig 2003-Lin et al. 1994

not identified DEN Roehrig 2003-Beasley&Askov, 2001

not identified DEN Roehrig 2003-Lok et al., 2001

not identified DEN Roehrig 2003

not identified DEN Roehrig 2003

not identified DEN Roehrig 2003

not identified DEN Roehrig 2003

not identified DEN Roehrig 2003

Page 2 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

not identified JE Roehrig 2003-Hasegawa et al., 1992

not identified JE Roehrig 2003-Cecilia & Gould, 1991

not identified JE Roehrig 2003-Morita et al., 2001

not identified JE Roehrig 2003

not identified JE Roehrig 2003

not identified JE Roehrig 2003

not identified LI Roehrig 2003-Gao et al., 1994

not identified LI Roehrig 2003-Jiang et al. 1993

not identified LI Roehrig 2003

not identified MVE Roehrig 2003-McMinn et al. 1995

not identified MVE Roehrig 2003

not identified MVE Roehrig 2003

not identified MVE Roehrig 2003

not identified MVE Roehrig 2003

not identified TBE Roehrig 2003-Holzmann et al. 1997, Mandl et al,

1989

not identified TBE Roehrig 2003

not identified TBE Roehrig 2003

not identified TBE Roehrig 2003

not identified TBE Roehrig 2003

not identified TBE Roehrig 2003

not identified TBE Roehrig 2003

not identified TBE Roehrig 2003

not identified TBE Roehrig 2003

not identified YF Roehrig 2003-Lobigs et al., 1987

not identified YF Roehrig 2003-Ryman et al, 1997

not identified YF Roehrig 2003

not identified YF Roehrig 2003

not identified YF Roehrig 2003

not identified DEN3 1047MP10 Lee et al. 1997







direct isolates DEN3 1153MP10 Lee et al. 1997

from patients



direct isolates DEN3 1239MP10 Lee et al. 1997

from patients



direct isolates DEN3 1047V1P10 Lee et al. 1997

from patients

direct isolates DEN3 1047V2P10 Lee et al. 1997

from patients



direct isolates DEN3 1153V1P10 Lee et al. 1997

from patients

direct isolates DEN3 1153V2P10 Lee et al. 1997

from patients

Page 3 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

Den3 1239V1P10 Lee et al. 1997







direct isolates DEN3 1239V2P10 Lee et al. 1997

from patients



direct isolates DEN3 1153CP5 Lee et al. 1997

from patients

direct isolates DEN3 1239CP5 Lee et al. 1997

from patients

direct isolates KUN NS2A1/MRM 61C Liu et al. 2004

from patients



D00246 KUN NS2A2/MRM 61C Liu et al. 2004





D00246 KUN NS5/MRM 61C Liu et al. 2004



D00246 rDEN4-2A-5 Hanley et al. 2003







AF326825 rDen4delta30-7129 Hanley et al. 2003









AF326827 rDEN4-7129 Hanley et al. 2003









AF326825 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

Page 4 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4 (could also be AF326825) Hanley et al. 2001

AF326573 rDEN4delta30-4995 Hanley et al. 2004







AF326827 rDEN4delta30-8092 Hanley et al. 2004







AF326827 rDEN4delta30-10634 Hanley et al. 2004







AF326827 rDEN4delta30-200-201 Hanley et al. 2004









AF326827 rDEN4delta30-436-437 Hanley et al. 2004







AF326827 rDEN4-4891 Blaney Jr. et al. 2003



AY648301 rDEN4-4995 Blaney Jr. et al. 2003



AY648301 rDEN4-7153 Blaney Jr. et al. 2003



AY648301 rDEN4-7162 Blaney Jr. et al. 2003



AY648301 rDEN4-7163 Blaney Jr. et al. 2003



AY648301 rDEN4-7182 Blaney Jr. et al. 2003



AY648301 rDEN4-7546 Blaney Jr. et al. 2003



AY648301 rDEN4-7630 Blaney Jr. et al. 2003



AY648301 rDEN4-10279 Blaney Jr. et al. 2003

AY648301 rDEN4-10275 Blaney Jr. et al. 2003

Page 5 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

AY648301 rDEN2/4delta30-4891 Blaney Jr. et al. 2003



rDEN2/4delta30-4995 Blaney Jr. et al. 2003

AY744147

rDEN2/4delta30-7153 Blaney Jr. et al. 2003

AY744147

rDEN2/4delta30-7162 Blaney Jr. et al. 2003

AY744147

rDEN2/4delta30-7163 Blaney Jr. et al. 2003

AY744147

rDEN2/4delta30-7182 Blaney Jr. et al. 2003

AY744147

rDEN2/4delta30-7546 Blaney Jr. et al. 2003

AY744147

rDEN2/4delta30-7630 Blaney Jr. et al. 2003

AY744147

rDEN2/4delta30-10279 Blaney Jr. et al. 2003

AY744147

rDEN2/4delta30-10275 Blaney Jr. et al. 2003

AY744147

JEVdelta39-44 Yoshio et al., 2004

JEVG42 Yoshio et al., 2004



JEV M4243 Yoshio et al., 2004









rDEN1mutF, derived from Western Pacific,74 Whitehead et al., 2003



rDEN1delta30, derived from Western Pacific,74 Whitehead et al., 2003









814669 DEN4 3'd 172-143 Men et al., 1996







814669 DEN4 3'd 172-113 Men et al., 1996







814669 DEN4 3'd 172-83 Men et al., 1996



814669 DEN4 3'd 243-183 Men et al., 1996

814669 DEN4 3'd 303-183 Men et al., 1996

Page 6 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

814669 DEN4 3'd 333-183 Men et al., 1996







814669 DEN4 3'd 384-183 Men et al., 1996







814669 DEN4 3'd 384-183/172-113 Men et al., 1996



814669 WNV, Kunjin strain Liu et al., 2006









NC_001563 Western TBE strain Neudoerfl c(delta28-43) – before Koffler et al, 2002

passaging

U27495 Western TBE strain Neudoerfl c(delta28-46) – after Koffler et al, 2002

passaging

U27495 Western TBE strain Neudoerfl c(delta28-48) – after Koffler et al, 2002

passaging

U27495 Western TBE strain Neudoerfl c(delta28-48/Q70L) Koffler et al, 2002







U27495 Western TBE strain Neudoerfl c(delta28-48/Du78-85) Koffler et al, 2002







U27495 Western TBE strain Neudoerfl c(delta28-48) – after Koffler et al, 2002

passaging

U27495 Western TBE strain Neudoerfl c(delta28-48) – after Koffler et al, 2002

passaging

U27495 Western TBE strain Neudoerfl c(delta28-48) – after Koffler et al, 2002

passaging

U27495 Western TBE strain Neudoerfl c(delta28-48) – after Koffler et al, 2002

passaging

U27495 Western TBE strain Neudoerfl c(delta28-48) – after Koffler et al, 2002

passaging

U27495 Western TBE strain Neudoerfl c(delta28-54) – after Koffler et al, 2002

passaging

U27495 Western TBE strain Neudoerfl c(delta28-54) – after Koffler et al, 2002

passaging

U27495 Western TBE strain Neudoerfl c(delta28-54) – after Koffler et al, 2002

passaging

U27495 Western TBE strain Neudoerfl c(delta28-57) – after Koffler et al, 2002

passaging

U27495 Western TBE strain Neudoerfl c(delta28-57) – after Koffler et al, 2002

passaging

U27495 Western TBE strain Neudoerfl c(delta28-54) – before Koffler et al, 2002

passaging

U27495 Western TBE strain Neudoerfl c(delta28-57) – before Koffler et al, 2002

passaging

Page 7 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

U27495 Western TBE strain Neudoerfl c(delta28-46) – before Koffler et al, 2002

passaging

U27495 Western TBE strain Neudoerfl c(delta28-48) – before Koffler et al, 2002

passaging

U27495 DEN2 6b2 escape mutant Lok et al., 2001



M29095/ New DEN2 6b2 passage control Lok et al., 2001

Guinea C

(Igarashi, 1978)

M29095/ New DEN2 10F2 escape mutant Lok et al., 2001

Guinea C

(Igarashi, 1978)

M29095/ New DEN2 10F2 passage control Lok et al., 2001

Guinea C

(Igarashi, 1978)

M29095/ New YF NS2Aalpha QST Kummerer and Rice, 2002

Guinea C

(Igarashi, 1978)

NC_002031 YF NS2Alpha QKV Kummerer and Rice, 2002

NC_002031 YF NS2Alpha QQT Kummerer and Rice, 2002

NC_002031 YF NS2Aalpha QET Kummerer and Rice, 2002

NC_002031 YF NS2Aalpha QRT Kummerer and Rice, 2002

NC_002031 YF NS2Aalpha QIT Kummerer and Rice, 2002

NC_002031 YF NS2Aalpha SKT Kummerer and Rice, 2002

NC_002031 YF NS2Aalpha RRS Kummerer and Rice, 2002



NC_002031 YF NS2Aalpha KLEEG reconstructed mutant Kummerer and Rice, 2002

NC_002031 YF NS2Aalpha KRET reconstructed mutant Kummerer and Rice, 2002

NC_002031 YF NS2Aalpha RRSTG reconstructed mutant Kummerer and Rice, 2002

NC_002031 YF NS2Aalpha SKT D343V double mutant Kummerer and Rice, 2002

NC_002031 YF NS2Aalpha SKT D343A double mutant Kummerer and Rice, 2002

NC_002031 YF NS2Aalpha SKT D343G double mutant Kummerer and Rice, 2002

NC_002031 YF NS3 D343V Kummerer and Rice, 2002

NC_002031 YF NS3 D343A Kummerer and Rice, 2002

NC_002031 YF NS3 D343G Kummerer and Rice, 2002

NC_002031 YF NS1 SB (YFiv5.2) Muylaert et al., 1996



YF NS1 G1a (YFiv5.2) Muylaert et al., 1996







NC_002031

YF NS1 G1b (YFiv5.2) Muylaert et al., 1996







NC_002031

YF NS1 G2a (YFiv5.2) Muylaert et al., 1996







NC_002031

Page 8 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

YF NS1 G2b (YFiv5.2) Muylaert et al., 1996







NC_002031

YF NS1 G1aG2a (YFiv5.2) Muylaert et al., 1996



NC_002031

YF NS1 G1bG2b (YFiv5.2) Muylaert et al., 1996



NC_002031

M29095 New pSV.NS2B/3 17-20 Matusan et al., 2001

Guinea C, Arias et

al. (1993), Teo &

Wright (1997),

Rice (1996),

Yusoff et al.

(2000)

M29095 New pSV.NS2B/3 32-36 Matusan et al., 2001

Guinea C, Arias et

al. (1993), Teo &

Wright (1997),

Rice (1996),

Yusoff et al.

(2000)

M29095 New pSVNS2B/3 63-66 Matusan et al., 2001

Guinea C, Arias et

al. (1993), Teo &

Wright (1997),

Rice (1996),

Yusoff et al.

(2000)

M29095 New pSVNS2B/3 95-96 Matusan et al., 2001

Guinea C, Arias et

al. (1993), Teo &

Wright (1997),

Rice (1996),

Yusoff et al.

(2000)

M29095 New pSVNS2B/3 179-181 Matusan et al., 2001

Guinea C, Arias et

al. (1993), Teo &

Wright (1997),

Rice (1996),

Yusoff et al.

(2000)

M29095 Dengue 2 NGC Kroshewski et al 2008



M29095 Dengue 2 NGC Kroshewski et al 2008

M29095 Dengue 2 NGC Kroshewski et al 2008

Page 9 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

M29095 Dengue 2 NGC Kroshewski et al 2008



M29095 Dengue 2 NGC Kroshewski et al 2008

M29095 Dengue 2 NGC Kroshewski et al 2008



M29095 Dengue 2 NGC Kroshewski et al 2008



M29095 Dengue 2 NGC Kroshewski et al 2008

M29095 Dengue 2 NGC Kroshewski et al 2008

M29095 Dengue 2 NGC Kroshewski et al 2008

M29095 Dengue 2 NGC Kroshewski et al 2008

M29095 Dengue 2 NGC Kroshewski et al 2008



M29095 Dengue 2 NGC Kroshewski et al 2008

M29095 Dengue 2 NGC Kroshewski et al 2008



M29095 Dengue 2 NGC Kroshewski et al 2008

M29095 WN pFLWNV 5'-3627 Zhang et al 2008



AF404756 WN pFLWNV 5'-3627 Zhang et al 2008



AF404756 WN pFLWNV 5'-3627 Zhang et al 2008



AF404756 WN pFLWNV 5'-3627 Zhang et al 2008



AF404756 WN pFLWNV 5'-3627 Zhang et al 2008



AF404756 WN pFLWNV 5'-3627 Zhang et al 2008



AF404756 WN pFLWNV 5'-3627 Zhang et al 2008

AF404756 WN pFLWNV 5'-3627 Zhang et al 2008



AF404756 WN pFLWNV 5'-3627 Zhang et al 2008



AF404756 WN pFLWNV 5'-3627 Zhang et al 2008



AF404756 WN pFLWNV 5'-3627 Zhang et al 2008



AF404756 D2-VEN90 Leitmeyer et al., 1999

AF100466 D2-VEN91 Leitmeyer et al., 1999

AF100150 D2-MEX95 Leitmeyer et al., 1999

AF100147 D2-MEX97 Leitmeyer et al., 1999

AF100146 D2-THAI96A Leitmeyer et al., 1999

AF100463 D2-THAI96B Leitmeyer et al., 1999

AF100464 D2-THAI95A Leitmeyer et al., 1999

AF100461 D2-THAI94A Leitmeyer et al., 1999

AF100459 D2-THAI94B Leitmeyer et al., 1999

AF100460 D2-THAI95B Leitmeyer et al., 1999

AF100462 D2-PR81 Leitmeyer et al., 1999

Page 10 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

AF100149 D2-COL86 Leitmeyer et al., 1999

AF100458 D2-MEX83 Leitmeyer et al., 1999

AF100151 D2-VEN90B Leitmeyer et al., 1999

AF100466 D2-VEN87 Leitmeyer et al., 1999

AF100465 D2-MEX92 Leitmeyer et al., 1999

AF100469 D2-MEX92B Leitmeyer et al., 1999

AF100148 D2-PERU95 Leitmeyer et al., 1999

AF100467 YF-17D Nickells and Chambers, 2003

X03700 KUN Leung et al 2008

AY274504 or KUN Leung et al 2008

D00246

AY274504 or KUN Leung et al 2008

D00246

AY274504 or KUN Leung et al 2008

D00246

AY274504 or KUN Leung et al 2008

D00246

AY274504 or KUN Leung et al 2008

D00246

AY274504 or KUN Leung et al 2008

D00246

AY274504 or KUN Leung et al 2008

D00246

U87411 DEN2 strain 16681 Alvarez et al 2008

U87411 DEN2 strain 16681 Alvarez et al 2008







U87411 DEN2 strain 16681 Alvarez et al 2008

U87411 DEN2 strain 16681 Alvarez et al 2008

U87411 DEN2 strain 16681 Alvarez et al 2008

U87411 DEN2 strain 16681 Alvarez et al 2008

U87411 DEN2 strain 16681 Alvarez et al 2008

U87411 DEN2 strain 16681 Alvarez et al 2008

U87411 DEN2 strain 16681 Alvarez et al 2008

U87411 DEN2 strain 16681 Alvarez et al 2008

U87411 DEN2 strain 16681 Alvarez et al 2008

U87411 DEN2 strain 16681 Alvarez et al 2008

U87411 DEN2 strain 16681 Alvarez et al 2008

U87411 DEN2 strain 16681 Alvarez et al 2008



U87411 DEN2 strain 16681 Alvarez et al 2008



U87411 DEN2 strain 16681 Alvarez et al 2008

U87411 DEN2 strain 16681 Alvarez et al 2008

U87411 DEN2 strain 16681 Alvarez et al 2008

U87411 DEN2 strain 16681 Alvarez et al 2008

U87411 DEN2 strain 16681 Alvarez et al 2008

U87411 DEN2 strain 16681 Alvarez et al 2008

Page 11 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

U87411 DEN2 strain 16681 Alvarez et al 2008

U87411 DEN2 strain 16681 Alvarez et al 2008

U87411 DEN2 strain 16681 Alvarez et al 2008

U87411 DEN2 strain 16681 Alvarez et al 2008

U87411 DEN2 strain 16681 Alvarez et al 2008

AB178040 rDEN 1-NIID02-20 Tajima et al 2008



AB178040 rDEN 1-NIID02-20 Tajima et al 2008

AB178040 rDEN 1-NIID02-20 Tajima et al 2008

AB178040 WN25A, WN25A.9 Chambers et al. 2008

AF404756 Rluc-Rep cDNA clone/pGEM-5NTR Zhang et al 2008



AF404756 Rluc-Rep cDNA clone/pGEM-5NTR Zhang et al 2008



AF404756 Rluc-Rep cDNA clone/pGEM-5NTR Zhang et al 2008



AF404756 Rluc-Rep cDNA clone/pGEM-5NTR Zhang et al 2008



AF404756 Rluc-Rep cDNA clone/pGEM-5NTR Zhang et al 2008



AF404756 Rluc-Rep cDNA clone/pGEM-5NTR Zhang et al 2008



AF404756 pFLWNV Zhang et al 2008



AF404756 pFLWNV Zhang et al 2008



AF404756 pFLWNV Zhang et al 2008



AF404756 pFLWNV Zhang et al 2008







AF404756 pFLWNV Zhang et al 2008



AF404756 pFLWNV Zhang et al 2008



AF196835 WNV Botha et al 2008

AF346318 WNV Botha et al 2008

EF429197 WNV Botha et al 2008

AB196923 JEVAT31strain-pMWJEATG1 Yamashiita et al 2008



AB196923 JEVAT31strain-pMWJEATG1 Yamashiita et al 2008



AB196923 JEVAT31strain-pMWJEATG1 Yamashiita et al 2008



AB196923 JEVAT31strain-pMWJEATG1 Yamashiita et al 2008



AB196923 JEVAT31strain-pMWJEATG1 Yamashiita et al 2008

Page 12 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

not identified pYF23 from cDNA clone of pACNR/FLYF Patkar and Kuhn 2008



not identified pYF23 from cDNA clone of pACNR/FLYF Patkar and Kuhn 2008







not identified pYF23 from cDNA clone of pACNR/FLYF Patkar and Kuhn 2008



not identified pYF23 from cDNA clone of pACNR/FLYF Patkar and Kuhn 2008









not identified pYF23 from cDNA clone of pACNR/FLYF Patkar and Kuhn 2008







not identified pYF23 from cDNA clone of pACNR/FLYF Patkar and Kuhn 2008



not identified pYF23 from cDNA clone of pACNR/FLYF Patkar and Kuhn 2008



AY648961 rDEN3-p3-7164 (includes adaption mut. V115A) Blaney Jr. et al. 2007



AY648961 rDEN3-p3-7164 (includes adaption mut. V115A) Blaney Jr. et al. 2007

AY648961 rDEN3-p3-7164 (includes adaption mut. V115A) Blaney Jr. et al. 2007

AY648961 rDEN3-p3-7164 (includes adaption mut. V115A) Blaney Jr. et al. 2007









AY648961 rDEN3-p3-7164 (includes adaption mut. V115A) Blaney Jr. et al. 2007



AY648961 rDEN3-p3-7164 (includes adaption mut. V115A) Blaney Jr. et al. 2007







AY648961 rDEN3-p3-7164 (includes adaption mut. V115A) Blaney Jr. et al. 2007



AY648961 rDEN3-p3-7164 (includes adaption mut. V115A) Blaney Jr. et al. 2007









AY648961 rDEN3-p3-7164 (includes adaption mut. V115A) Blaney Jr. et al. 2007



AY648961 rDEN3-p3-7164 (includes adaption mut. V115A) Blaney Jr. et al. 2007







AY648961 rDEN3-p3-7164 (includes adaption mut. V115A) Blaney Jr. et al. 2007







not identified WNR-CNS1-5luc replicon Suzuki et al 2008

Page 13 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

not identified WNR-CNS1-5luc replicon Suzuki et al 2008

not identified WNR-CNS1-5luc replicon Suzuki et al 2008

not identified WNR-CNS1-5luc replicon Suzuki et al 2008

not identified WNR-CNS1-5luc replicon Suzuki et al 2008



not identified WNR-CNS1-5luc replicon Suzuki et al 2008



not identified WNR-CNS1-5luc replicon Suzuki et al 2008

not identified WNR-CNS1-5luc replicon Suzuki et al 2008



not identified WNR-CNS1-5luc replicon Suzuki et al 2008

not identified WNR-CNS1-5luc replicon Suzuki et al 2008

not identified WNR-CNS1-5luc replicon Suzuki et al 2008

not identified WNR-CNS1-5luc replicon Suzuki et al 2008



not identified WNR-CNS1-5luc replicon Suzuki et al 2008

not identified WNR-CNS1-5luc replicon Suzuki et al 2008

not identified WNR-CNS1-5 no luc replicon Suzuki et al 2008



not identified WNV genome Suzuki et al 2008



not identified WNR-CNS1-5luc replicon Suzuki et al 2008



not identified WNR-CNS1-5luc no luc replicon Suzuki et al 2008



AY303793-4 JE-CJN Chiou and Chen 2007

AY303793-4 JE-CJN Chiou and Chen 2007







AY303793-4 JE-CH1392 Chiou and Chen 2006

AF254451 JE-CH1392 Chiou and Chen 2007

AF254452 JE-T1P1 Chiou and Chen 2007



AY303791-2 or JE-T1P1 Chiou and Chen 2007

AF254453



AY303791-2 or JE-CC27 Chiou and Chen 2007

AF254453

AY303795-8 JE-CC27 Chiou and Chen 2007

AY303795-8 JE-CC27 Chiou and Chen 2007









AY303795-8 JE-CC27 Chiou and Chen 2007







AY303795-8 DV2 8 strains Wong et al 2007

Page 14 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

AJ556803–AJ556 DV2 8 strains Wong et al 2007

814

AJ556803–AJ556 DV2 8 strains Wong et al 2007

814

AJ556803–AJ556 DV2 8 strains Wong et al 2007

814

AJ556803–AJ556 TBE pTNd/c (replicon) Hoenninger et al 2008

814

U27495 TBE pTNd/c (replicon) Hoenninger et al 2008



U27495 TBE pTNd/c (replicon) Hoenninger et al 2008



U27495 TBE pTNd/c (replicon) Hoenninger et al 2008



U27495 TBE pTNd/c (replicon) Hoenninger et al 2008







U27495 TBE pTNd/c (replicon) Hoenninger et al 2008



U27495 TBE pTNd/c (replicon) Hoenninger et al 2008

U27495 TBE pTNd/c (replicon-9a) Hoenninger et al 2008



U27495 TBE pTNd/c (replicon-∆polyA) Hoenninger et al 2008



U27495 TBE pTNd/c (replicon-∆var) Hoenninger et al 2008



U27495 IRES-eGFP (Gehrke et. al 2005) (cassette) Hoenninger et al 2008







U27495 IRES-ME (Orlinger et. al 2006) (cassette) Hoenninger et al 2008







U27495 IRES-a771c and IRES-26a(Orlinger et. al 2007) Hoenninger et al 2008

(cassette)



U27495 DV2 Strain 16681 and its plasmid strain16681pst Sangiambut et al 2008







M84727,U87411, DV2 Strain 16681 and its plasmid strain16681pst Sangiambut et al 2008

or NC_001474







M84727,U87411, DV2 Strain 16681 and its plasmid strain16681pst Sangiambut et al 2008

or NC_001474



M84727,U87411, DV2 Strain 16681 and its plasmid strain16681pst Sangiambut et al 2008

or NC_001474

Page 15 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

M84727,U87411, DV2 Strain 16681 and its plasmid strain16681pst Sangiambut et al 2008

or NC_001474

M84727,U87411, DV4 Strain 814669 FRhL clone Lai et al 2007

or NC_001474







AF326573 DV4 Strain 814669 C6/36 clone Lai et al 2007









AF326573 TBE - pTd/bc Orlinger et al 2007



U27495 TBE - pTd/bc Orlinger et al 2007



U27495 TBE - pTd/bc Orlinger et al 2007







U27495 TBE - pTd/bc Orlinger et al 2007



U27495 TBE - pTd/bc Orlinger et al 2007







U27495 WNV replicon Rossi et al 2007







not identified WNV replicon Rossi et al 2007

not identified WNV replicon Rossi et al 2007

not identified WNV replicon Rossi et al. 2007









not identified WNV replicon Rossi et al. 2007









not identified WNV replicon Rossi et al. 2007









not identified WNV Evans and Seeger 2007

Page 16 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

not identified WNV replicon Evans and Seeger 2007









not identified WNII genome Evans and Seeger 2007









not identified WNV Jia et. Al 2007







EF657887 DV1 Bordignon et al 2007

AF226687

Page 17 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

Mutation (protein name) Natural/Artificial

Env-K307E Artificial ESM caused by nucleotid change in Egene



Env-K307E Artificial ESM caused by nucleotid change in Egene



NS1 E1114G Artificial M

NS2A M11581I Artificial M

NS2A T1321A Artificial M

NS3 L1602F Artificial M

NS3 S1632P Artificial M

NS3 D1865G Artificial M

NS5 N2583I Artificial M

NS5 E2664G Artificial M

NS5 I3362T Artificial M

NS5 K2924R Artificial M

E Protein Asp181Tyr Domain I Artificial ESM



E Protein Ala123Lys Domain II Artificial ESM

E Protein Gly368Arg Domain III Artificial ESM

E Protein L68P or F, E Protein S154N, E Protein Glycosylation at E-154-156 Natural Mutation – Early cell culture passage of WNI



E Protein S154N, Glycosylation NYS E-154-156 Natural Mutation – Recovered from mouse brain inoc w/ WNI-568

E Protein L68P, Y155N, Glycosylation at 155-157 NST Natural Mutation – mosquito passed strain of WNI

E Protein Y155N, K307E Glycosylation at E 155-157 NST Artificial ESM of WNI-25

E: V35A, K386N, R486K Artificial ESM



E: S169P ,G275R,F43K Artificial ESM



E Protein Lys388Asn, Asp390Asn Natural Mutation – obtained from clone of DEN2 Mexican strain



E Protein Asp390Asn Natural Mutation – obtained from clone of DEN2 Mexican strain



E Protein Asp390His Natural Mutation – obtained from clone of DEN2 Mexican strain

E Protein Asp390Asn Natural Mutation – obtained from clone of DEN2 Mexican strain



E Protein Asp390Asn Natural Mutation – obtained from clone of DEN2 Mexican strain



E Protein Asp390Asn Natural Mutation – obtained from clone of DEN2 Mexican strain



E Protein Asp390His Natural Mutation – obtained from clone of DEN2 Mexican strain

E Protein Asp390His Natural Mutation – obtained from clone of DEN2 Mexican strain

E Protein T69I Artificial ESM via Mab selecting

E Protein E71D Artificial ESM via Mab selecting

E Protein S112G Artificial ESM via Mab selecting

E Protein I124N Artificial ESM via Mab selecting

E Protein F279S Artificial ESM via Mab selecting

E Protein T293I Artificial ESM via Mab selecting

E Protein K307E Artificial ESM via Mab selecting

E Protein E311G Artificial ESM via Mab selecting

Page 18 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

E Protein Q52K or R Artificial ESM via Mab selecting

E Protein I126T Artificial ESM via Mab selecting

E Protein K136E Artificial ESM via Mab selecting

E Protein I270S Artificial ESM via Mab selecting

E Protein S275P Artificial ESM via Mab selecting

E Protein G333D Artificial ESM via Mab selecting

E Protein D308N Artificial ESM via Mab selecting

E Protein S310P Artificial ESM via Mab selecting

E Protein K311Q or N Artificial ESM via Mab selecting

E Protein A126E Artificial ESM via Mab selecting

E Protein R128S Artificial ESM via Mab selecting

E Protein F274V Artificial ESM via Mab selecting

E Protein S276R Artificial ESM via Mab selecting

silent nuc NS4B CA7129-7130UU Artificial ESM via Mab selecting

E Protein D67G Artificial ESM via Mab selecting



E Protein A71V Artificial ESM via Mab selecting

E Protein A123K Artificial ESM via Mab selecting

E Protein K171E Artificial ESM via Mab selecting

E Protein D181Y Artificial ESM via Mab selecting

E Protein Q233K Artificial ESM via Mab selecting

E Protein G368R Artificial ESM via Mab selecting

E Protein Y384H Artificial ESM via Mab selecting

E Protein S389R Artificial ESM via Mab selecting

E Protein N71K, Y, or H Artificial ESM via Mab selecting

E Protein D72G Artificial ESM via Mab selecting

E Protein M125I Artificial ESM via Mab selecting

E Protein D155G Artificial ESM via Mab selecting

E Protein T158I Artificial ESM via Mab selecting

E Protein Ala18Ser, Ala54Glu Artificial M via passaging through mouse brain







E Protein Phe277Ser Artificial M via passaging through mouse brain







E Protein Glu401Lys, Thr403Ile Artificial M via passaging through mouse brain







E Protein Thr266Ile, Ile268Thr Artificial M via passaging through vero cells



E Protein Phe191Val Artificial M via passaging through vero cells







E Protein Phe191Leu, Glu291Val Artificial M via passaging through vero cells



prM Lys26Gln, E Protein Phe191Leu, Lys202Arg, Ile268Val Artificial M via passaging through vero cells

Page 19 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

prM Lys26Thr, E Protein Ile268Ser Artificial M via passaging through vero cells







E Protein Phe191Leu, Lys202Arg Artificial M via passaging through vero cells







E Protein Thr155Ala; loss glycosylation site at E-133 Artificial M via passaging through Ae. albopictus cells



E Protein Thr155Met; loss glycosylation site at E-133 Artificial M via passaging through Ae. albopictus cells



NS2A Ala30Pro Artificial M via cloning replicon RNA w/ puromycin resistance, luciferase and

beta-galactosidase gene



NS2A Asn101Asp Artificial M via cloning replicon RNA w/ puromycin resistance, luciferase and

beta-galactosidase gene



NS5 Pro270Ser Artificial M via cloning replicon RNA w/ puromycin resistance, luciferase and

beta-galactosidase gene

NS4B Pro101Leu, Nuc C7129, silent nuc in NS4B C7359U, silent nuc in NS5 Artificial M via 5-Fu mutatgenesis of DEN4 virus

U9047C





NS4B Pro101Leu, NS4B nuc C7124U, silent nuc NS4B CA7129-7130UU and Artificial M via cloning Pro101Leu into rDEN4delta30 vaccine candidate

deletion in 3'-UTR from nuc 10478-10507







NS4B Pro101Leu, NS4B nuc C7124U, silent nuc NS4B U6521C and silent nuc Artificial M via cloning Pro101Leu into wt DEN4 cDNA

NS4B CA7129-7130UU







NS5 DR21-22AA Artificial M via Kunkel mutagenesis

NS5 RK22-23AA Artificial M via Kunkel mutagenesis

NS5 KE23-24AA Artificial M via Kunkel mutagenesis

NS5 EE26-27AA Artificial M via Kunkel mutagenesis

NS5 KD46-47AA Artificial M via Kunkel mutagenesis

NS5 EE157-158AA Artificial M via Kunkel mutagenesis

NS5 KH200-201AA Artificial M via Kunkel mutagenesis

NS5 RH246-247AA Artificial M via Kunkel mutagenesis

NS5 EK253-254AA Artificial M via Kunkel mutagenesis

NS5 KE356-357AA Artificial M via Kunkel mutagenesis

NS5 KK387-388AA Artificial M via Kunkel mutagenesis

NS5 KK388-389AA Artificial M via Kunkel mutagenesis

NS5 RE396-397AA Artificial M via Kunkel mutagenesis

NS5 EE397-398AA Artificial M via Kunkel mutagenesis

NS5 DK436-437AA Artificial M via Kunkel mutagenesis

NS5 RE500-501AA Artificial M via Kunkel mutagenesis

NS5 EE520-521AA Artificial M via Kunkel mutagenesis

NS5 DK523-524AA Artificial M via Kunkel mutagenesis

NS5 KK524-525AA Artificial M via Kunkel mutagenesis

NS5 KD525-526AA Artificial M via Kunkel mutagenesis

Page 20 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

NS5 KD596-597AA Artificial M via Kunkel mutagenesis

NS5 KE641-642AA Artificial M via Kunkel mutagenesis

NS5 ER642-643AA Artificial M via Kunkel mutagenesis

NS5 EK645-646AA Artificial M via Kunkel mutagenesis

NS5 KE649-650AA Artificial M via Kunkel mutagenesis

NS5 DR654-655AA Artificial M via Kunkel mutagenesis

NS5 RE750-751AA Artificial M via Kunkel mutagenesis

NS5 ED808-809AA Artificial M via Kunkel mutagenesis

NS5 ED820-821AA Artificial M via Kunkel mutagenesis

NS5 DK827-828AA Artificial M via Kunkel mutagenesis

NS5 KE877-878AA Artificial M via Kunkel mutagenesis

NS5 EE878-879AA Artificial M via Kunkel mutagenesis

NS5 KE23-24AA, KH200-201AA Artificial M via Kunkel mutagenesis

NS5 KE23-24AA, RE396-397AA Artificial M via Kunkel mutagenesis

NS3 S68D, nuc U4725G, NS3 S68D, nuc C4726A, deletion in 3'-UTR from nuc Artificial M, charge-to-alanine and point mutations previously done by site-

10478-10507 directed mutagenesis; chimeric clone created via cloning techniques



E Protein D225A, nuc A1612C, NS4B T102A, nuc A7131G, deletion in 3'-UTR Artificial M, charge-to-alanine and point mutations previously done by site-

from nuc 10478-10507 directed mutagenesis; chimeric clone created via cloning techniques



NS4B M48L, nuc A6969U, NS4B G119R, nuc G7182C, deletion in 3'-UTR from Artificial M, charge-to-alanine and point mutations previously done by site-

nuc 10478-10507 directed mutagenesis; chimeric clone created via cloning techniques



NS3 G103R, nuc G4830A, NS5 V1821, nuc G8106A, deletion in 3'-UTR from Artificial M, charge-to-alanine and point mutations previously done by site-

nuc 10478-10507 directed mutagenesis; chimeric clone created via cloning techniques







NS4B L112F, nuc A7163C, deletion in 3'-UTR from nuc 10478-10507 Artificial M, charge-to-alanine and point mutations previously done by site-

directed mutagenesis; chimeric clone created via cloning techniques



NS3 I1597T Artificial M via site directed mutagenesis



NS3 S1632P Artificial M via site directed mutagenesis



NS4B V2351A Artificial M via site directed mutagenesis



NS4B L2354S Artificial M via site directed mutagenesis



NS4B L2354F Artificial M via site directed mutagenesis



NS4B G2361S Artificial M via site directed mutagenesis



NS4B A2482V Artificial M via site directed mutagenesis



NS5 K2510R Artificial M via site directed mutagenesis



3'-UTR A > U Artificial M via site directed mutagenesis

3'-UTR A > U Artificial M via site directed mutagenesis

Page 21 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

NS3 I1597T, DEN2 structural genes, 3'-UTR deletion 10478-10507 Artificial M via cloning of previously site directed mutants into rDEN2/4delta30

chimeric virus

NS3 S1632P, DEN2 structural genes, 3'-UTR deletion 10478-10507 Artificial M via cloning of previously site directed mutants into rDEN2/4delta30

chimeric virus

NS4B V2351A, DEN2 structural genes, 3'-UTR deletion 10478-10507 Artificial M via cloning of previously site directed mutants into rDEN2/4delta30

chimeric virus

NS4B L2354S, DEN2 structural genes, 3'-UTR deletion 10478-10507 Artificial M via cloning of previously site directed mutants into rDEN2/4delta30

chimeric virus

NS4B L2354F, DEN2 structural genes, 3'-UTR deletion 10478-10507 Artificial M via cloning of previously site directed mutants into rDEN2/4delta30

chimeric virus

NS4B G2361S, DEN2 structural genes, 3'-UTR deletion 10478-10507 Artificial M via cloning of previously site directed mutants into rDEN2/4delta30

chimeric virus

NS4B A2482V, DEN2 structural genes, 3'-UTR deletion 10478-10507 Artificial M via cloning of previously site directed mutants into rDEN2/4delta30

chimeric virus

NS5 K2510R, DEN2 structural genes, 3'-UTR deletion 10478-10507 Artificial M via cloning of previously site directed mutants into rDEN2/4delta30

chimeric virus

3'-UTR A > U, DEN2 structural genes, 3'-UTR deletion 10478-10507 Artificial M via cloning of previously site directed mutants into rDEN2/4delta30

chimeric virus

3'-UTR A > U, DEN2 structural genes, 3'-UTR deletion 10478-10507 Artificial M via cloning of previously site directed mutants into rDEN2/4delta30

chimeric virus

Core Protein deletion from residues 39-44 Artificial M

Core Protein Gly42Ala Artificial M



Core Protein Gly42A, Pro43A Artificial M









incidental: prM T173M, nuc C612U, purposeful: 3'-SL sequence (nucleotide Artificial M generated by recombination between linearized prs424/DEN1 WP

mutations): deletion of A @ 7 and U @ 73 and U4A and C74U and pcr frament containing mutations

purposeful: 3'UTR deletion of nucleotides from 172 to 143, incidental: E protein Artificial M generated by recombination of pR424DEN1WP and linearized

T552S, nuc A1748U PCR product









deletion of 3'NC region from nucleotides 172-143 artificial introduced deletion mutations







deletion of 3'NC region from nucleotides 172-113 artificial introduced deletion mutations







deletion of 3'NC region from nucleotides 172-83 artificial introduced deletion mutations



deletion of 3'NC region from nucleotides 243-183 artificial introduced deletion mutations

deletion of 3'NC region from nucleotides 303-183 artificial introduced deletion mutations

Page 22 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

deletion of 3'NC region from nucleotides 333-183 artificial introduced deletion mutations







deletion of 3'NC region from nucleotides 384-183 artificial introduced deletion mutations







deletion of 3'NC region from nucleotides 172-113 and 384-183 artificial introduced deletion mutations



NS2A A30P Artifical M, substitution









Protein C deletion from AA 28-43 Artificial mutation – deletion via cloning



Protein C deletion from AA 28-46, P57L (nuc C302U) Artificial mutation – deletion via cloning



Protein C deletion from AA 28-48, Du78-85 (nuc Du364-387) Artificial mutation – deletion via cloning



Protein C deletion from AA 28-48, point mutation Q70L Artificial mutation – deletion cloning, point mutation via passaging







Protein C deletion from AA 28-48, duplication from I78 to L85 Artificial mutation – deletion via cloning, duplication via passaging







Protein C deletion from AA 28-48, T81M (nuc C374U) Artificial mutation – deletion cloning, duplication mutation via passaging



Protein C deletion from AA 28-48, Du59-K77 (nuc Du302-363) Artificial mutation – deletion cloning, point mutation via passaging



Protein C deletion from AA 28-48, T72I (nuc C347 U) Artificial mutation – deletion cloning, point mutation via passaging



Protein C deletion from AA 28-48, N56I, V66F, T72I (nuc A299U, G328U, Artificial mutation – deletion cloning, point mutation via passaging

C347U)

Protein C deletion from AA 28-48, V66F, T81M (nuc G328U, C374U) Artificial mutation – deletion cloning, point mutation via passaging



Protein C deletion from AA 28-54, K79I (nuc A368U) Artificial mutation – deletion cloning, point mutation via passaging



Protein C deletion from AA 28-54, T81M (nuc C374U) Artificial mutation – deletion cloning, point mutation via passaging



Protein C deletion from AA 28-54, Q90L (nuc A401U) Artificial mutation – deletion cloning, point mutation via passaging



Protein C deletion from AA 28-57, Duplications 24-27 and 58-78 (nuc Artificial mutation – deletion cloning, duplication mutation via passaging

Duplications 202-213 and 304-366)

Protein C deletion from AA 28-57, K79I (nuc A368U) Artificial mutation – deletion cloning, point mutation via passaging



Protein C deletion from AA 28-54 Artificial mutation – deletion via cloning



Protein C deletion from AA 28-57 Artificial mutation – deletion via cloning

Page 23 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

Protein C deletion from AA 28-46 Artificial mutation – deletion via cloning



Protein C deletion from AA 28-48 Artificial mutation – deletion via cloning



prm G15S, E E311G Artificial ESM – passage in BHK-21 in presence of DEN2 specific Mab 6B2



E Protein P53S, E71D, S112G, I124N, F402F/L Artificial M – passage control







prM G15S, E Protein T69I, E71D, S112G, I124N, F402L Artificial ESM – passage in BHK-21 in presence of DEN2 specific Mab 10F2







E Protein P53S, S112G, I124N, F402L Artificial M – passage control







NS2A/B cleavage site, QKT->QST (AAG->TCG) Artifical M, substitution via cloning







NS2A/B cleavage site, QKT->QKV (ACT->GTT) Artificial M, substitution

NS2A/B cleavage site, QKT->QQT (AAG->CAG) Artificial M, substitution

NS2A/B cleavage site, QKT->QET (AAG->GAG) Artifical M, substitution

NS2A/B cleavage site, QKT->QRT (AAG->AGG) Artifical M, substitution

NS2A/B cleavage site, QKT->QIT (AAG->ATC) Artifical M, substitution

NS2A/B cleavage site, QKT->SKT (CAG->TCG) Artifical M, substitution

NS2A/B cleavage site, QKT->RRS (CAG-AAG-ACT->AGG-AGG-AGT) Artifical M, substitution



NS2A/B cleavage site, QST->QKLEEGST Artifical M, substitution

NS2A/B cleavage site, QIT->QKRETIT Artifical M, substitution

NS2A/B cleavage site, QST->QRRSTGST Artifical M, substitution

NS2A/B cleavage site, QKT->SKT, NS3 D343V Artifical M, substitution

NS2A/B cleavage site, QKT->SKT , NS3 D343A Artifical M, substitution

NS2A/B cleavage site, QKT->SKT, NS3 D343G Artifical M, substitution

NS3 D343V Artifical M, substitution

NS3 D343A Artifical M, substitution

NS3 D343G Artifical M, substitution

NS1 silent mutation, no AA changes; nuc: AAGC2845-2848CTCA, T3076C, Artifical M, oligonucleotide-directed mutagenesis, substitution

G3079T, A3082C

NS1 S132A, AGC2846-2848GCT Artifical M, oligonucleotide-directed mutagenesis, substitution









NS1 N130A, AA2840-2841GC Artifical M, oligonucleotide-directed mutagenesis, substitution









NS1 T210A, A3080G, A3082C Artifical M, oligonucleotide-directed mutagenesis, substitution

Page 24 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

NS1 N208A, AA3074-3075GC Artifical M, oligonucleotide-directed mutagenesis, substitution









NS1 S132A, T210A, AGC2846-2848GCT, A3080G, A3082C Artifical M, oligonucleotide-directed mutagenesis, substitution







NS1 N130A, N208A, AA2840-2841GC, AA3074-3075GC Artifical M, oligonucleotide-directed mutagenesis, substitution







NS3: E17A/E19A/D20A Artifical M, charge to alanine substitution









NS3 G32A/ Y33A Artifical M, charge to alanine substitution









NS3 K63A/ R64A/E66A, Artifical M, charge to alanine substitution









NS3 V95A/Q96A Artifical M, charge to alanine substitution









NS3 E179A/D180A/ D181A Artifical M, charge to alanine substitution









NS5 methyltransferase: S56T, S56A Artificial mutations



NS5 methyltransferase: W87I Artificial mutations

NS5 methyltransferase: W87K Artificial mutations

Page 25 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

NS5 methyltransferase: W87Y Artificial mutations



NS5 methyltransferase: D131A Artificial mutations

NS5 methyltransferase: D131E Artificial mutations



NS5 methyltransferase: D131N Artificial mutations



NS5 methyltransferase: K76A/D79A Artificial mutations

NS5 methyltransferase: D79A/L80I Artificial mutations

NS5 methyltransferase: G81A/G83A/G85A Artificial mutations

NS5 methyltransferase: D146A/E149A Artificial mutations

NS5 methyltransferase: G48A, G48P Artificial mutations



NS5 methyltransferase: K46A/R47A/E49A Artificial mutations

NS5 methyltransferase: E138A/K139A/D141A Artificial mutations



NS5 methyltransferase: E192A/K193A/E195A Artificial mutations

NS5 methyltransferase: K61T Artificial mutations



NS5 methyltransferase: K61Q Artificial mutations



NS5 methyltransferase: D146S Artificial mutations



NS5 methyltransferase: D146L Artificial mutations



NS5 methyltransferase: D146P Artificial mutations



NS5 methyltransferase: D146R Artificial mutations



NS5 methyltransferase: D146T Artificial mutations

NS5 RNA dependent RNA polymerase: W751R Artificial mutations



NS1: P138L Artificial mutations



5' UTR: U38-ins Artificial mutations



5' UTR: G35U Artificial mutations

Page 26 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls









Env- domain III (residues 305, 326, and 380) Neuroadapted viruses compared

NS2A: I59N artificial

NS2A: T149P artificial



NS2A: I59R artificial



NS2A: I59V artificial



NS2A: K198S artificial



NS2A: I59Y artificial



NS2A: I59S artificial



NS2A: I59T artificial



NS5 artificial

5' A artificial







5'-3' A artificial

5' B artificial

5'-3' B artificial

5' UAR 1 artificial

5' UAR 2 artificial

5' UAR 3 artificial

5' UAR 4 artificial

5' UAR 5 artificial

5' UAR 6 artificial

5' UAR 7 artificial

3' UAR 1 artificial

3' UAR 2 artificial



3' UAR 3 artificial



3' UAR 4 artificial

3' UAR 5 artificial

3' UAR 6 artificial

3' UAR 7 artificial

5'-3' UAR 1 artificial

5'-3' UAR 2 artificial

Page 27 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

5'-3' UAR 3 artificial

5'-3' UAR 4 artificial

5'-3' UAR 5 artificial

5'-3' UAR 6 artificial

5'-3' UAR 7 artificial

NS1: N130A artificial



NS1: N130A/N207A artificial

NS1: N207A artificial

E: Y155N, Y155S artificial

5' UAR 1 (replicon) artificial



5' UAR 2 (replicon) artificial



5' UAR 3 (replicon) artificial



5' UAR 4 (replicon) artificial



5' UAR 5 (replicon) artificial



5' UAR 6 (replicon) artificial



5' UAR 1 (genome) artificial



5' UAR 2 (genome) artificial



5' UAR 3 (genome) artificial



5' UAR 4 (genome) artificial







5' UAR 5 (genome) artificial



5' UAR 6 (genome) artificial



NS4B: C102S natural

NS4B: E249G natural

NS4B: A79T natural

Walker A motif of Helicase: G199A, K200A, T201A artificial



Walker A motif of Helicase: K200R, K200Q, K200N, K200D, K200E, K200H artificial



Motif IV: R461A, R464A artificial



Motif IV: Q457A,R458A artificial



Motif IV: R459A, G460A,V462A, and G463A artificial

Page 28 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

NS3: Q341A/ T342A/D343A, D343V artificial



NS3: E338A/D339A, I334A/P345A/S346A, H353A/D354A artificial







NS3: E347A/P348A/W349A, W349A/N350A artificial



NS3: E347A, P348A,W349A, N350A artificial









NS3: W349F, W349Y, W349H artificial







NS3: W349Q, W349L, W349K artificial



SIN-NS2B3: S138A, R461Q artificial



3'UTR (already include deleted nucleotides 173-143) 50 deletions artificial



3'UTR (already include deleted nucleotides 173-143) 61 deletions artificial

3'UTR (already include deleted nucleotides 173-143) 80 deletions artificial

3'UTR (already include deleted nucleotides 173-143) 86 deletions artificial









3'UTR (already include deleted nucleotides 173-143) 116A deletions artificial



3'UTR (already include deleted nucleotides 173-143) 116B deletions artificial







3'UTR (already include deleted nucleotides 173-143) 146 deletions artificial



3'UTR (already include deleted nucleotides 173-143)/ 31 deletions artificial









3'UTR 31 deletions artificial



NS5-3' : rDEN3-3'UTR-D4 artificial







NS5-3' : rDEN3-3'UTR-D4(with 30nt deletions) artificial







5'CS and 3'CS in C protein coding region all A-U switched to U-A artificial

Page 29 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

5'CS and 3'CS in C protein coding region all G-C switched to C-G artificial

5'CS and 3'CS in C protein coding region all A-U switched to G-C artificial

5'CS and 3'CS in C protein coding region reverse sequences artificial

5'CS and 3'CS in C protein coding region reverse complimentary sequences artificial



single-nucleotide substitutions in CS both 5' and 3' regions (terminal ends), m1 artificial

and m2

single-nucleotide substitutions in CS both 5' and 3' regions, m4 and m7 artificial

single-nucleotide substitutions in CS both 5' and 3' regions, m3,m8,m10,m11 artificial



single-nucleotide substitutions in CS both 5' and 3' regions, m5, m6 artificial

single-nucleotide substitutions in CS both 5' and 3' regions, m9 artificial

5' UTR m1 insertion artificial

5' UTR m2 insertion artificial



3' UTR m1 insertion artificial

3' UTR m2 insertion artificial

double-nucleotide substitutions in CS both 5' and 3' regions,m4 and m7 artificial

combination

double-nucleotide substitutions in CS both 5' and 3' regions,m4 and m7 artificial

combination in whole viral genome

5'CS double-nucleotide substitution of m4/m7, 3'CS double-nucleotide artificial

substitution of m4/m7

5'CS double-nucleotide substitution of m4/m7, 3'CS double-nucleotide artificial

substitution of m4/m8

E138glu/ NS4A162leu (L1variant) natural - substitution occurred in transfection

E138lys/NS4A162Ile (S1variant) natural - substitution occurred in transfection







E306glu (L3variant) natural - substitution occurred in transfection

E306lys (S5variant) natural - substitution occurred in transfection

E306glu (L4variant) natural - substitution occurred in transfection



E306lys (S1variant) natural - substitution occurred in transfection







E36asn/E135ser/E389asp (L1variant) natural - substitution occurred in transfection



E36ser/E135ser/E389asp (L3variant) natural - substitution occurred in transfection

E36ser/E135ser/E389asn (S6variant) natural - substitution occurred in transfection









E36asn/E135Ile/E389asp (S8variant) natural - substitution occurred in transfection







E: G385A natural- one outbreak to another

Page 30 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

NS1: nucleotide changes natural- one outbreak to another



NS1: nucleotide changes natural- one outbreak to another



NS1: A815G natural- one outbreak to another



C17rluc reporter (first 17 aa of the capsid plus renilla luciferase, rest of prM and artificial

E except for TM2 deleted)

C27rluc reporter (first 27 aa of the capsid plus renilla luciferase, rest of prM and artificial

E except for TM2 deleted)

C37rluc reporter (first 37 aa of the capsid plus renilla luciferase, rest of prM and artificial

E except for TM2 deleted)

C17fluc reporter (first 17 aa of the capsid plus fire fly luciferase, rest of prM and artificial

E except for TM2 deleted)

C17fluc-FMDV2A reporter (first 17 aa of the capsid plus firefly luciferase, rest of artificial

prM and E except for TM2 deleted) 2A from foot and mouth disease



C17fluc-TaV2A reporter (first 17 aa of the capsid plus firefly luciferase, rest of artificial

prM and E except for TM2 deleted) 2A from Thosea asigna

NS5:GDD-GAA artificial

all but 9 adenines of 49 removed from poly A tract of 3' NCR artificial



all adenines of 49 removed from poly A tract of 3' NCR artificial



entire variable region removed of 3' NCR artificial



enhanced green fluorescent protein under control of an EMCV IRES cloned into artificial

C17fluc-TaV2A



encodes for the TBEV prM and E proteins under control of an EMCV IRES artificial

cloned into C17fluc-TaV2A



A771C substitution in oligo(A) loop/insertion of 19 additional adenines in the artificial

same oligo(A) loop cloned into C17fluc-TaV2A



C:K6A/K7A artificial







C:K73A/K74A artificial









C:R85A/K86A artificial







C:R97A/R98A artificial

Page 31 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

C:R85A/K86A, R97A/R98A artificial



DENV4v3 A-G mutation at nucleotide 1458 natural - 3 cycles of neutralization and propagation









DENV4v4 C-T mutation at nucleotide 1465 natural - 3 cycles of neutralization and propagation









pTBEV-bc was created from Taking TBEV and removing the prM and E regions artificial

and inserting with a EMCV IRES in the 3'NCR - E: D308A, A317T

mutant clones of TBEV-bc E: D308A/A317T natural after passage of pTBEV-bc



mutation in IRES : consistent with nucleotide 771 of the EMCV genome A-C natural after passage of pTBEV-bc







mutation in IRES:consistent with nucleotide 771 of the EMCV genome A-C and natural after passage of pTBEV-bc

E: D308A/A317T

mutation in IRES : U769A/poly(A), caused insertion of multiple Adenines natural after passage of pTBEV-bc

(between 11-35)



NS3: 117K artificial







NS4B: E249G artificial

NS5: P528H artificial

NS2A: D73H artificial









NS2A: M108K artificial









NS2A: A30P artificial









NS4B: E34A/K35A,E45A/E46A, R74A/K75A, E121A/K123A,D35A/R37A, artificial

E132A/R135A, E163A/R164A

Page 32 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

NS4B: E22A/K24A artificial









NS4B: E22A/K24A artificial









NS2A: V61A, prM: P54S natural-variant in cell culture







E: T276P natural after passage in cell culture

Page 33 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

Phenotype

rendered antibody G8D11unable to bind to the glycoprotein E in

radioimmunoprecipitation therefore allowing for replication

rendered antibody G8D11unable to bind to the glycoprotein E in

radioimmunoprecipitation therefore allowing for replication

attenuation & temperature sens.

attenuation & temperature sens.

attenuation & temperature sens.

attenuation & temperature sens.

attenuation & temperature sens.

attenuation & temperature sens.

attenuation & temperature sens.

attenuation & temperature sens.

attenuation & temperature sens.

temperature sens. NOT attenuated

no diff from wild in ts or plaque size. was attenuated.more sens. to ph

(conform stab)

smaller plaques, not ts. attenuated. more sens. to ph (conform stab)

ts, normal plaque. attenuated. more sens. to ph (conform stab)

Neuroinvasive and Neurovirulent



Neuroinvasive and Neurovirulent

Loss Neuroinvasiveness, still neurovirulent

Loss Neuroinvasiveness, still Neurovirulent





Less fusion of cells, failure to agglutinate erythrocytes, smaller plaques



Produced small lytic plaque, Medium neurovirulence, var. mortality (10 to 7th viral

titer(increased from parental strain); decreased pH fusion threshhold -0.2



P6-P10, severe hindlimb paralysis; 100% mortality; >10 to 7th viral

titer(increased from parental strain); decreased pH fusion threshhold -0.7



P6-P10, severe hindlimb paralysis; 100% mortality; >10 to 7th viral

titer(increased from parental strain); decreased pH fusion threshhold -0.6 to -

0.8

P6-10 had 0.5-2 x 10 to 7th viral titers (10-100 fold higher); larger forming

plaques earlier in time(3 days as opposed to 6 days)

P6-10 had 0.5-2 x 10 to 7th viral titers (10-100 fold higher); larger forming

plaques earlier in time(3 days as opposed to 6 days); decreased pH fusion

threshhold -0.8 to -1.0

P6-10 had 0.5-2 x 10 to 7th viral titers (10-100 fold higher); larger forming

plaques earlier in time(3 days as opposed to 6 days)

P6-10 had 0.5-2 x 10 to 7th viral titers (10-100 fold higher); larger forming

plaques earlier in time(3 days as opposed to 6 days)

Page 35 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

P6-10 had 0.5-2 x 10 to 7th viral titers (10-100 fold higher); larger forming

plaques earlier in time(3 days as opposed to 6 days); decreased pH fusion

threshhold -0.6 to -0.8

P6-10 had 0.5-2 x 10 to 7th viral titers (10-100 fold higher); larger forming

plaques earlier in time(3 days as opposed to 6 days); decreased pH fusion

threshhold -0.6 to -0.8

smaller plaque produced at 7 days; extensive syncytium formation at 5-6

days; increased pH fusion threshhold +0.4

smaller plaque produced at 7 days; extensive syncytium formation at 5-6

days; increased pH fusion threshhold +0.4

no change in initiation of RNA synthesis (72% of wt) in BHK, 17 fold increase

in RNA in BHK, 50x increase in colonies in human(HEK293/HEp-2); did not

inhibit IFN-beta driven transcription

decrease in initiation of RNA synthesis (55% of wt) in BHK, 13 fold increase

in RNA synthesis in BHK; 20x increase in hunan(HEK293/HEp-2)



decrease in initiation of RNA synthesis (12% of wt) in BHK, 1.8 fold increase

in RNA synthesis in BHK

decreased plaque size in C6/36 cells but increased plaque size in Vero/HuH7

cells; low percentage of total infection in Ae. agyptis and no dissemination; no

infection in mosquito heads; not ts in vero/HuH7 or decreased replication in

mice; increased rep w/ SCID/HuH7 mice

smaller plaque size in C6/36, increased plaque size in Vero/HuH7 cells; high

percentage total infection but no dissemination; no ts in Vero/HuH7 and no

change in replication in mice; mild decrease in replication in SCID/HuH7 mice



small plaque size in C6/36, increased plaque size in Vero/HuH7; low total

infection and no dissemination; decreased infection in head to 25% in

Toxorhynchites splendens; no ts in Vero/HuH7, no change in replication in

mice; increased replication in SCID/HuH7 mice

not ts, no attenuation

ts in Vero & HuH7, attenuated replication in suckling mice

ts in HuH7, attenuated replication in suckling mice

ts in Vero & HuH7, no attenuation

not ts, no attenuation

ts in HuH7, attenuated replication in suckling mice

ts in HuH7

not ts, no attenuation

not ts, no attenuation

ts in HuH7, attenuated replication in suckling mice

ts in Vero & HuH7, attenuated replication in suckling mice

ts in Vero & HuH7, no attenuation

ts in HuH7, no attenuation

ts in Vero & HuH7, no attenuation

ts in Vero & HuH7, attenuated replication in suckling mice

ts in Vero & HuH7, no attenuation

ts in Vero & HuH7, no attenuation

ts in HuH7, no attenuation

not ts, attenuated replication in suckling mice

not ts, attenuated replication in suckling mice

Page 36 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

ts in Vero & HuH7, no attenuation

not ts, no attenuation

ts in Vero & HuH7, attenuated replication in suckling mice

ts in Vero , no attenuation

not ts, no attenuation

ts in Vero & HuH7, attenuated replication in suckling mice

not ts, no attenuation

ts in Vero & HuH7, attenuated replication in suckling mice

ts in Vero & HuH7, no attenuation

ts in HuH7, attenuated replication in suckling mice

not ts, attenuated replication in suckling mice

not ts, attenuated replication in suckling mice

ts in Vero & HuH7, no attenuation

ts in Vero & HuH7, attenuated replication in suckling mice

significant decrease in viral titer in SCID/HuH7 mice, slight decrease in viral

titer in rhesus monkeys, protected monkey against DEN4 challenge



slight decrease in viral titer in SCID/HuH7, significant decrease in viral titer in

rhesus monkeys, protected monkey against DEN4 challenge



mild decrease in viral titer in SCID/HuH7, mild decrease in viral titer in rhesus

monkeys, protected monkey against DEN4 challenge



significant decrease in viral titer in SCID/HuH7 mice compared to delta30

alone, no decrease compared to charge-to-alanine, significant decrease in

viral titer in rhesus monkeys, protected monkey against DEN4 challenge



significant decrease in viral titer in SCID/HuH7 mice compared to delta30 &

charge-to-alanine, significant decrease in viral titer in rhesus monkeys,

protected monkey against DEN4 challenge

increased plaque size in Vero cells, attenuated replication in mouse brain



increased plaque size in Vero cells, attenuated replication in mouse brain



increased plaque size in Vero cells, no attenuation of replication in mouse

brain

increased plaque size in Vero cells, no attenuation of replication in mouse

brain

increased plaque size in Vero cells, no attenuation of replication in mouse

brain

increased plaque size in Vero cells, no attenuation of replication in mouse

brain

increased plaque size in Vero cells, no attenuation of replication in mouse

brain

increased plaque size in Vero cells, no attenuation of replication in mouse

brain

no increase in plaque size in Vero cells

no increase in plaque size in Vero cells

Page 37 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

substantial increase in replication in Vero cells



substantial increase in replication in Vero cells



substantial increase in replication in Vero cells



substantial increase in replication in Vero cells



substantial increase in replication in Vero cells



substantial increase in replication in Vero cells



substantial increase in replication in Vero cells



substantial increase in replication in Vero cells



substantial increase in replication in Vero cells



substantial increase in replication in Vero cells



core protein only localized in cytoplasm, was not present in nucleus

core protein did not colocalize with nucleolin, distributed as filamentous

strands in nucleus, found in cytoplasm also

core protein was not transported into nucleus, only in cytoplasm, smaller

infectious plaque, decreased release of infectious particles, increased release

of defective particles, impaired RNA replication in Vero cells, earlier peak of

protein synthesis, imaired neuroinvasiveness



mild decrease in viral titers in rhesus monkeys, lower Ab titers than wt, but

still generated protective immunity

less than 1 day viremic and low viral titers (0.8 log 10 PFU), lower Ab titers

relative to wt but still generated protective immunity, decreased viral

replication by 100 fold in HuH-7 mice, highly infective in T. splendens

(approach log of 1.0 concentration after intrathoracic injection)



1-10 faint plaques, plaques similar to wt DEN4, 9 mm(C6/36 cells), late and

faint plaque production(LLC-MK2 cells @ 9 days), reduced Ab titers relative

to wt

1-10 faint plaques, plaques similar to wt DEN4, 10 mm(C6/36 cells), no

plaque @ 6 days, faint plaques at 9 days(LLC-MK2 cells), high Ab titers

against protein E

no plaques produced(LLC-MK2 or C6/36), no RNA present (lethal mutation)



not studied due to mutations not being stable after cell passage

20-30 faint plaques, smaller plaque than wt, 6 mm(C6/36 cells), late and faint

plaque production(LLC-MK2 cells), high Ab titers against protein E and NS1

Page 38 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

RNA present via IFA, smaller plaque than wt, 5mm(C6/36 cells), late and faint

plaque production, present at 9 days(LLC-MK2 cells), poor Ab titer response



no plaques produced(LLC-MK2), RNA present via IFA, smaller plaque than

wt, 3mm(C6/36 cells), no plaque(LLC-MK2 cells), poor Ab titer response



no plaques produced(LLC-MK2), RNA present via IFA, smaller plaque than

wt, 3mm(C6/36 cells)

higher IFN-beta produced in Hep2 cells infected w/ SFV, abortive replication

of WNV-Kun in A549 cells, smaller plaque foci on A549 cells, decreased viral

titers in A549 cells, more IFN-alpha/beta produced in mice, higly attenuated

in mice(lower virulence), protected from challenge w/ WNV-NY99



infection of BHK-21 w/o mouse passaging; infection similar to wt



increased plaque size in BHK-21 cells



restored ability of virus to infect BHK-21 cells



infection of BHK-21 w/o mouse passaging, reduced and delayed release of

viral particles in corneal endothelial cells, atten index (LD/ID) > 10 to 5 in

mice

infection of BHK-21 w/o mouse passaging, reduced and delayed release of

viral particles in corneal endothelial cells, atten index (LD/ID) > 10 to 5 in

mice

restored ability of virus to infect BHK-21 cells



restored ability of virus to infect BHK-21 cells



restored ability of virus to infect BHK-21 cells



restored ability of virus to infect BHK-21 cells



restored ability of virus to infect BHK-21 cells



restored ability of virus to infect BHK-21 cells



restored ability of virus to infect BHK-21 cells



restored ability of virus to infect BHK-21 cells



restored ability of virus to infect BHK-21 cells



restored ability of virus to infect BHK-21 cells



inability of virus to infect BHK-21 cells



inability of virus to infect BHK-21 cells

Page 39 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

reduced ability of virus to infect BHK-21 cells



inability of virus to infect BHK-21 cells



lower absorbance, lower fusion, decrease in titer due to 42 C of approx 3

PFU/ml

lower absorbance, normal fusion, decrease in titer due to 42C of approx 4

PFU/ml



decrease in absorbance, normal fusion, decrease in titer due to 42C by

approx 5 PFU/ml



decrease in absorbance, normal fusion, decrease in titer due to 42C by

approx 3 PFU/ml



no plaque produced, no infectious particles produced (lacks capsid and RNA,

still has E and protein), plaque production restored with trans NS1-2A



small plaque formation with infectivity similar to wild type

no plaque formation

no plaque formation

plaque formation and infectivity similar to wild type

no plaque formation

no plaque formation

plaque formation and infectivity similar to wild type, cleavage of NS2Aalpha

more efficient than wild type

plaque formation and infectivity similar to wild type

plaque formation and infectivity similar to wild type

smaller plaque formation, infectivity similar to wild type

plaque formation with infectivity similar to wild type

plaque formation and infectivity similar to wild type

plaque formation and infectivity similar to wild type

larger plaque formation than wild type, infectivity similar to wild type

larger plaque formation than wild type, infectivity similar to wild type

larger plaque formation than wild type, infectivity similar to wild type

phenotypically identical to wild type



small plaque, lower virus yield, delayed cytopathological effects, faster

migration in gel than wt, slight decrease in protein production, reduced RNA

production up to 24 hrs postinf., attenuation in mice (16% mortality with mild

illnesses in survivors)

small plaque, lower virus yield, delayed cytopathological effects, faster

migration in gel than wt, slight decrease in protein production, reduced RNA

production up to 24 hrs postinf., attenuation in mice (16% mortality with mild

illnesses in survivors)

medium plaque(smaller than wt), wt level viral yields, cytopathological effects

similar to wt(24-48hrs postinf), faster migration than wt in gel, similar levels of

RNA accumulation in wt, similar virulence and phenotype in mice

Page 40 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

medium plaque(smaller than wt), wt level viral yields, cytopathological effects

similar to wt(24-48hrs postinf), faster migration than wt in gel, similar levels of

RNA accumulation in wt, similar virulence and phenotype in mice



tiny plaque, lower viral yields, slower migration than wt on gel, lower protein

production, reduced RNA production up to 24 hrs postinf, attenuation in

mice(no deaths and minimal signs of disease)

tiny plaque, lower viral yields, slower migration than wt on gel, lower protein

production, reduced RNA production up to 24 hrs postinf, attenuation in

mice(no deaths and minimal signs of disease)

interaction between NS2B/NS3 retained, no severe inhibition of proteinase

activity









interaction between NS2B/NS3 retained, low titres recovered (ts), probable

restriction of replication









interaction between NS2B/NS3 retained, no severe inhibition of proteinase

activity, reduced plaque size, titer comprable to wild type, initial lag in virus

release, no temp. sensitivity observed









reduction of intereaction between NS2B/NS3 observed, low titres recovered

(ts),probable restriction of replication









interaction between NS2B/NS3 retained, no severe inhibition of proteinase

activity, reduced plaque size, titer comprable to wild type, initial lag in virus

release, no temp. sensitivity observed









Both changes cause loss of N7 metransferase, S56T has 100% 2' Ome

activity. No growth.

Loss of both N7 metransferase and 2' Ome activity. No Growth.

Loss of both N7 metransferase and 2' Ome activity. No Growth.

Page 41 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

Loss of both N7 metransferase and 2' Ome activity. Temperature sensitive

growth.

Loss of both N7 metransferase and 2' Ome activity. No Growth.

100 % N7 metransferase activity and loss of 2' Ome activity. Temperature

sensitive growth.

Loss of both N7 metransferase and 2' Ome activity. Temperature sensitive

growth.

Loss of both N7 metransferase and 2' Ome activity. No Growth.

Loss of both N7 metransferase and 2' Ome activity. No Growth.

Loss of both N7 metransferase and 2' Ome activity. No Growth.

Loss of both N7 metransferase and 2' Ome activity. No Growth.

Loss of both N7 metransferase and 2' Ome activity. G48A has temperatures

sensitive growth, G48P no growth.

loss of 2' Ome activity. Temperature sensitive growth.

enhanced activity in both N7metransferase and 2' Ome. enhanced growth at

higher temperatures.

enhanced activity in 2'Ome. Growth only at lower temperature.

contributes to the rescue of viral replication of D146S mutation by improving

N7 methylation activity

contributes to the rescue of viral replication of D146S mutation by improving

N7 methylation activity

produces virus, some N7 methylation activity retained, no 2' Ome activity



produces no virus, no N7 methylation activity retained, no 2' Ome activity



produces no virus, no N7 methylation activity retained, no 2' Ome activity



produces no virus, no N7 methylation activity retained, no 2' Ome activity



some N7 methylation activity retained, no 2' Ome activity

contributes to the rescue of viral replication of D146S mutation, improves

polymerase activity

does not contribute to the rescue of viral replication of D146S mutation



contributes to the rescue of viral replication of D146S mutation, do not

increase cap methylations

contributes to the rescue of viral replication of D146S mutation, do not

increase cap methylations

Page 42 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls









Determined these 3 residues were critical for neuroinvasiveness

loss of RNA replication, low VLP production with reversion

rescues the defect in I59N (compensatory), allows for virus

assembly/secretion

severely impaired RNA replication, low VLP production with reversion to I59Y



enhanced RNA replication, no effect on VLP production



loss of RNA replication, no effect on VLP production



enhanced RNA replication, no effect on VLP production



enhanced RNA replication, no effect on VLP production



enhanced RNA replication, no effect on VLP production



reduced luciferase activity, RNA synthesis probably impaired

luciferase activity similar to WT, suggests efficient synthesis, however this is

time sensitive, after 96 h. activity is severly reduced and no replication occurs



luciferase activity similar to WT, suggests efficient synthesis

reduced luciferase activity, no RNA synthesis

reduced luciferase activity, no RNA synthesis

maintains base pairs, able to replicate

disrupted hybridization, reduced RNA replication

hybridization occurred, mismatch present, replication still occurred

disrupted hybridization, reduced RNA replication

disrupted hybridization, reduced RNA replication

disrupted hybridization, reduced RNA replication

no replication detected

hybridization disrupted, no RNA replication

generated stem loop mismatch,disrupted hybridization, reduced RNA

replication

hybridization occurred, mismatch present in stem loop , replication still

occurred

efficient RNA replication

reduced RNA replication

reduced RNA replication

no replication detected

restores replication capability defected in 3'UAR mut.

restored some RNA replication lost in 5' and 3' UAR mut., reduction of

synthesis probably due to a mismatch at position -8

Page 43 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

high levels of RNA replication

high levels of RNA replication

high levels of RNA replication

high levels of RNA replication

no replication detected

no infectious viral particles found after one passage blocks the transition of

WT In trans.

no infectious viral particles found after one passage

infectious viral particles found after one passage

may destabilize doman I/II interface leading to attenuation

forms stable duplex with 3'UAR,viral translation occurs, replication occurs



forms stable duplex with 3'UAR,viral translation occurs, replication occurs



forms stable duplex with 3'UAR,viral translation occurs, replication occurs



does not form stable duplex with 3'UAR, viral translation occurs, replication

does not occur

forms stable duplex with 3'UAR,viral translation occurs, replication does not

occur

forms stable duplex with 3'UAR,viral translation occurs, replication occurs



formed p.f.u. ,exhibited immunofluorescence, no adaptive change in

transfected virus

formed p.f.u. ,exhibited immunofluorescence, no adaptive change in

transfected virus

formed p.f.u. ,exhibited immunofluorescence, no adaptive change in

transfected virus

no p.f.u. formed, exhibited some delayed immunofluorescence may indicate

reversion,adaptation made which led to rescue of replication in replicon



formed pinpoint p.f.u.,exhibited delayed immunofluorescence, adaptation

increased efficiency of replication and infectivity

formed pinpoint p.f.u. exhibited delayed immunofluorescence, no adaptive

change in transfected virus

temperature sens. At 41°C, attenuation in mice

reduced RNA synthesis in host cells

may alter the hydrophobicity of NS4B

all lost ATPase and helicase activity, no infectious virus recovered upon

transfection

all lost ATPase and helicase activity



complete loss of ATPase activity and helicase activity,no infectious virus

recovered upon transfection

loss of most ATPase activity and helicase activity,no infectious virus

recovered upon transfection

ATPase and helicase activity conserved, G460A produced viable infectious

virus

Page 44 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

produced WT plaques 72 h.p.t., retained WT luciferase activity, retained

packaging efficiency

produced small plaques 72 h.p.t.,loss of luciferase activity except for

H353A/D354A which retained WT levels, reduced packaging efficiency in all



produced no plaques 72 h.p.t., retained WT luciferase activity,no packaging

efficiency observed

all but W349A produced plaques at 72 h.p.t., all retained WT luciferase

activity, packaging efficiency observed in all but W349A, W349A blocks

production of infectious virus particles, but not the non-infectious particles



all but W349H produced plaques, W349H produced no plauqes, all retained

WT luciferase activity, packaging efficiency was conserved in all but W349H



All produced no plaques, all retained WT luciferase activity, no packaging

efficiency was conserved

previously known to inhibit enzymatic activity, did not effect -trans

complementation in W349A, enzymatic activity deemed unneeded

recovered in cDNA clones in C6/36cells ,viable, but replication restricted in

Vero cells

recovered in cDNA clones in C6/36cells

recovered in cDNA clones in C6/36cells

recovered in cDNA clones in C6/36cells , restricted in replication in SCID-

HuH-7 mice more than 10 fold, in Vero replicated like WT, In C6/36 replicated

10 fold lower than WT,in vero contained a coding change in M, and a single

nt substitution in the 3' UTR, also a mixed pop. found at NS5 stop that coded

for 2 extra amino acids, not detected in rhesus, but did infect





recovered in cDNA clones in C6/36cells ,viable, but replication restricted in

Vero cells

recovered in cDNA clones in C6/36cells ,deemed unstable due to natural

mutation of 8nt deletion and a single substitution of A→G at nt265



recovered in cDNA clones in C6/36cells ,viable, but replication restricted in

Vero cells

recovered in cDNA clones in C6/36cells , restricted in replication in SCID-

HuH-7 mice a bit less than 10 fold , in Vero replicated like WT, no replication

in C6/36 cells, in vero contained a single NS4B coding change, not detected

in rhesus but did infect, infects Toxorynchites but is attenuated



recovered in cDNA clones in C6/36cells , also recovered in Vero cells,

deemed unstable due to natural mutaion of 25nt deletion

in vero replicated nearly 100 fold higher than WT, in C6/36 WT replication

observed, detected in 75% of inoculated rhesus monkeys (suggests that ∆30

is necessary for attenuation

restricted in replication in SCID-HuH-7 mice more than 10 fold, in Vero

replicated like WT, in C6/36 replicatd 10 fold lower than WT, in vero

contained a 3'UTR substitution, not detected in rhesus but did infect

no luciferase activity at 48h post transfection

Page 45 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

no luciferase activity at 48h post transfection

no luciferase activity at 48h post transfection

no luciferase activity at 48h post transfection

no luciferase activity at 48h post transfection



showed drastic reduction of RNA replication



no detectable reduction in RNA replication (WT)

showed greater than 50% reduction in RNA replication



showed about a 20% reduction in RNA replication

showed about a 30% reduction in RNA replication

showed drastic reduction of RNA replication

replicated much better than m2, suggests A at position 2 in 3'CS is critical for

RNA replication

showed drastic reduction of RNA replication

showed drastic reduction of RNA replication

no detectable reduction in RNA replication (WT), no interference with genome

packaging, no reduction in foci forming effectiveness

no detectable defect despite 5' and 3' CS mutations and amino acid changes



no detectable RNA replication at 48 hours



10,000 times less efficent at producing foci fomation than WT at 48h, but 4

day incubation produced viable replicons

virus growth faster than the S1 variant

growth titer increased after one passage greater than that of LP growth

signifying that Neuro-2a cells prefer small plaque variants, more efficient

inhibition with heparin, survival times longer in mice

higher neurovirulence than the S5 variant



higher neurovirulence than the S1 variant, virus growth faster than S1 variant



growth titer increased after one passage greater than that of LP growth

signifying that Neuro-2a cells prefer small plaque variants, more efficient

inhibition with heparin

higher neurovirulence than S6/S8 variants



higher neurovirulence than S6/S8 variants

growth titer increased after one passage greater than that of LP growth

signifying that Neuro-2a cells prefer small plaque variants, more efficient

inhibition with heparin, slower virus growth than LP variants, lowest growth

titer of four variants

growth titer increased after one passage greater than that of LP growth

signifying that Neuro-2a cells prefer small plaque variants, survival times

longer in mice, slower virus growth than LP variants

caused a change in the viral genome of the second outbreak V129I,

happened at B-cell reactive regions, this could change the presentation of the

epitope producing different antibodies

Page 46 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

caused T349A, happened at B-cell reactive regions



caused T351A, happened at B-cell reactive regions



caused a change in the viral genome of the second outbreak K272R,

happened at B-cell reactive regions

initial input RNA translated , no replication in BHK cells, no detectable NS1

protein

initial input RNA translated, then decrease in activity followed by increase

again (biphasic)

initial input RNA translated, then decrease in activity followed by increase

again (biphasic)

initial input RNA translated, then decrease in activity followed by increase

again (biphasic)

activity highter than in C17fluc, suggests more efficient release of fluc at the

2A site versus 2B site in C17fluc



activity highter than in C17fluc-FMDV2A, suggests more efficient release of

fluc at the 2A site versus 2B site in C17fluc

abolishes viral RNA replication, but not translation

did not significantly affect the efficiency of cap-dependent translation, does

not affect RNA replication

did not significantly affect the efficiency of cap-dependent translation, does

not affect RNA replication

slight decrease in efficiency of cap-dependent translation, does not effect

RNA replication

no strong affect on the efficiency of cap-dependent translation, eventually

displayed reduced luciferase activity than parental replicon, replication delay



no strong affect on the efficiency of cap-dependent translation, eventually

displayed reduced luciferase activity than parental replicon, replication delay



did not demonstrate significanly increased efficiency of input RNA, increase

as compared to IRES ME suggesting positive effect on RNA replication

efficiency

initial high nucleolar staining, but large decrease after 72 h, nuclear

localization affected by time only in PS cells, reduced replication in C6/36

gained additional substitution mutation A6V in mammal cells

staining reduced significantly from parent cells, but appeared in 80%+ cells,

no increase in nucleoplasmic staining, found less frequently in Vero cells,

reduced replication in C6/36, gained additional point mutation at C317A

resulting in a A73N and A74N in later days

staining reduced significantly from parent cells, but appeared in 80%+ cells,

no increase in nucleoplasmic staining, found less frequently in Vero cells,

reduced replication in C6/36

initial high nucleolar staining, but large decrease after 72 h, nuclear

localization affected by time only in PS cells, not viable in vero cells, reduced

replication in C6/36

Page 47 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

not viable



resulted in E174K substitution in the E protein, could not be neutralized by

IgG5H2∆D plaque growth on vero cells similar to FRhL parent strain ,

mutation did not hinder growth , substitution reduced antibody binding affinity



resulted in L176P substitution in the E protein , somewhat neutralized by

IgG5H2∆D, plaque growth on vero cells similar to C6/36 parent strain,

mutation did not hinder growth, substitution reduced antibody binding affinity



virus is viable but replicates much slower than WT



produced more infectious particles than original bc, better specific infectivity

than original bc

produced more infectious particles than original bc, produced more RNA than

original BC but less than wildtype, greater specific infectivity than original bc



produced more infectious particles than original bc, titer equal to that of WT,

greatest increase in specific infectivity

still functional, efficiency varies with respect to number of adenines,

moderate increase in expression from the cap-dependent cistron, no strong

effect on E protein expression

noncytopathic, High Colony Formation efficiency, reduced replication and

decreased staining , reverted to WT when introduced to a NS1-5 ET2AN

replicon or genome

exhibited WT phenotype

exhibited WT phenotype

100% colony-forming efficiency, antigen staining less intense than WT

suggesting reduced WNR replication, weak expression of eGFP fluroescence

, infected Huh7 cells appeared healthy and dividing but were antigen positive,

produced smaller antigen positive foci than WT in vero cells, very little CPE

after 72hpi, not virulent in mice, IFN response delayed and deceased

compared to WT

100% colony-forming efficiency, antigen staining less intense than WT

suggesting reduced WNR replication, weak expression of eGFP fluroescence

,produced smaller antigen positive foci than WT in vero cells, very little CPE

after 72hpi, not virulent in mice

3.5% colony-forming efficiency, WT antigen staining efficiency, WT eGFP

fluroescence intensity, infected Huh7 cells appeared moribund like WT and

were antigen positive, produced somewhat smaller antigen postive foci than

WT but larger than other two NS2A mutations, CPE readily evident after

72hpi, virulent to mice and showed no reversion to WT





did not show competent RNA replication

Page 48 aaa8a56d-9ebb-4ce6-8e8e-af81155136db.xls

greater than 95% postive for beta-lactamase showed replication, produced

colonies after 3 weeks of selection in G418, demonstrated WT NS4B

replication and translation these results were not cell specific,IFN- signaling

pathway proved to remain functional in host cell , allowed for more stat1

phosphorylation than WT but overall stat1 protein was decreased from

parental cells, allowed for some induction of ISG





greater than 95% postive for beta-lactamase showed replication, produced

colonies after 3 weeks of selection in G418, demonstrated WT NS4B

replication and translation these results were not cell specific, IFN stimulation

completely inhibited, completely blocked STAT1 phosphorylation,caused less

cell death than WT

smaller plaques,somewhat ts at 41C, attenuative in mice, lower replication

efficiency, lower infection rate of C. pipens, may have a defect blocking cell-

cell spread, reversion occurred in vivo

extra information







Virus name Reference Protein/Residue/Region Location

DEN2 Wong et al 2007 E: 129 Domain II

DEN2 Wong et al 2007 NSI n/a

TBEV Hoenninger et al 2008 Internal Poly (A) tract 3' NCR

TBEV Hoenninger et al 2008 TM2 E protein

TBEV Hoenninger et al 2008 mutations IRES

DV2 Sangiambut et al 2008 C n/a

TBEV Orlinger et al 2007 E/ IRES n/a

TBEV Orlinger et al 2008 E: E387K n/a

TBEV Orlinger et al 2009 E: D308A n/a

WNV Rossi et al 2007 NS2A n/a

WNV/KUNV Rossi et al 2007 NS2A n/a









Page 49

extra information







Associated Amino Acid

usually either I or V , ( I for DV1, V for DV3, and L for DV4)

n/a

n/a

n/a

n/a

n/a

n/a

Glutamate

Asparagine

n/a

A30P mutation









Page 50

extra information







Significance/Important Information

important for type specific neutralizing antibodies, amino acid specific probably for importance of E protein function

stimulates cytotoxic T-Cell responses

no impact on translation or replication in mammal cells when removed or truncated

serves as an internal signal sequence for establishing the proper topology of the polyprotein in the ER membrane

as previously hypothesized, mutations causing weaker binding of the ribosome do not increase the fitness of bicistronic RNA

nucleolar localization not needed for dengue virus replication

mutations in these reason seemed to be sole reason for titer increases after passage

when aa is changed, result is alteration of surface charge

when aa is changed, result is alteration of surface charge, increases affinity of virus binding to cell surface

important for the establishment of noncytopathic persistent replication in mammalian cells

not the same between strains of North American/Australian variety, likely unsuitable for an attenuated vaccine strain









Page 51

extra information









tance of E protein function





tein in the ER membrane

crease the fitness of bicistronic RNA







ing to cell surface



an attenuated vaccine strain









Page 52

NS2A







D00246 KUN Liu et al. 2004 NS2A Ala30Pro

NS2A1/MRM

61C



D00246 KUN Liu et al. 2004 NS2A

NS2A2/MRM Asn101Asp

61C



NC_001563 WNV, Kunjin Liu et al., 2006 NS2A A30P

strain









NC_002031 YF NS2Alpha Kummerer and NS2A/B

QKV Rice, 2002 cleavage site,

QKT->QKV

(ACT->GTT)

NC_002031 YF NS2Alpha Kummerer and NS2A/B

QQT Rice, 2002 cleavage site,

QKT->QQT

(AAG->CAG)

NC_002031 YF NS2Aalpha Kummerer and NS2A/B

QET Rice, 2002 cleavage site,

QKT->QET

(AAG->GAG)

NC_002031 YF NS2Aalpha Kummerer and NS2A/B

QRT Rice, 2002 cleavage site,

QKT->QRT

(AAG->AGG)

NC_002031 YF NS2Aalpha Kummerer and NS2A/B

QIT Rice, 2002 cleavage site,

QKT->QIT (AAG-

>ATC)

NC_002031 YF NS2Aalpha Kummerer and NS2A/B

SKT Rice, 2002 cleavage site,

QKT->SKT

(CAG->TCG)

NC_002031 YF NS2Aalpha Kummerer and NS2A/B

RRS Rice, 2002 cleavage site,

QKT->RRS

(CAG-AAG-ACT-

>AGG-AGG-

AGT)

NC_002031 YF NS2Aalpha Kummerer and NS2A/B

KLEEG Rice, 2002 cleavage site,

reconstructed QST-

mutant >QKLEEGST

NC_002031 YF NS2Aalpha Kummerer and NS2A/B

KRET Rice, 2002 cleavage site,

reconstructed QIT->QKRETIT

mutant

NC_002031 YF NS2Aalpha Kummerer and NS2A/B

RRSTG Rice, 2002 cleavage site,

reconstructed QST-

mutant >QRRSTGST

NC_002031 YF NS2Aalpha Kummerer and NS2A/B

SKT D343V Rice, 2002 cleavage site,

double mutant QKT->SKT, NS3

D343V









Page 53

NS2A







NC_002031 YF NS2Aalpha Kummerer and NS2A/B

SKT D343A Rice, 2002 cleavage site,

double mutant QKT->SKT ,

NS3 D343A

NC_002031 YF NS2Aalpha Kummerer and NS2A/B

SKT D343G Rice, 2002 cleavage site,

double mutant QKT->SKT, NS3

D343G

AY274504 or KUN Leung et al 2008 NS2A: I59N

D00246

AY274504 or KUN Leung et al 2008 NS2A: T149P

D00246

AY274504 or KUN Leung et al 2008 NS2A: I59R

D00246

AY274504 or KUN Leung et al 2008 NS2A: I59V

D00246

AY274504 or KUN Leung et al 2008 NS2A: K198S

D00246

AY274504 or KUN Leung et al 2008 NS2A: I59Y

D00246

AY274504 or KUN Leung et al 2008 NS2A: I59S

D00246

AY274504 or KUN Leung et al 2008 NS2A: I59T

D00246

not identified WNV Rossi et al. 2007 NS2A: D73H









not identified WNV Rossi et al. 2007 NS2A: M108K









not identified WNV Rossi et al. 2007 NS2A: A30P









Page 54

NS2A







Artificial M via cloning replicon RNA w/ puromycin resistance,

luciferase and beta-galactosidase gene





Artificial M via cloning replicon RNA w/ puromycin resistance,

luciferase and beta-galactosidase gene





Artifical M, substitution









Artificial M, substitution









Artificial M, substitution









Artifical M, substitution









Artifical M, substitution









Artifical M, substitution









Artifical M, substitution









Artifical M, substitution









Artifical M, substitution









Artifical M, substitution









Artifical M, substitution









Artifical M, substitution









Page 55

NS2A







Artifical M, substitution









Artifical M, substitution









artificial



artificial



artificial



artificial



artificial



artificial



artificial



artificial



artificial









artificial









artificial









Page 56

NS2A







no change in initiation of RNA synthesis (72% of wt) in BHK, 17 fold increase

in RNA in BHK, 50x increase in colonies in human(HEK293/HEp-2); did not

inhibit IFN-beta driven transcription



decrease in initiation of RNA synthesis (55% of wt) in BHK, 13 fold increase in

RNA synthesis in BHK; 20x increase in hunan(HEK293/HEp-2)





higher IFN-beta produced in Hep2 cells infected w/ SFV, abortive replication

of WNV-Kun in A549 cells, smaller plaque foci on A549 cells, decreased viral

titers in A549 cells, more IFN-alpha/beta produced in mice, higly attenuated in

mice(lower virulence), protected from challenge w/ WNV-NY99





small plaque formation with infectivity similar to wild type









no plaque formation









no plaque formation









plaque formation and infectivity similar to wild type









no plaque formation









no plaque formation









plaque formation and infectivity similar to wild type, cleavage of NS2Aalpha

more efficient than wild type









plaque formation and infectivity similar to wild type









plaque formation and infectivity similar to wild type









smaller plaque formation, infectivity similar to wild type









plaque formation with infectivity similar to wild type









Page 57

NS2A







plaque formation and infectivity similar to wild type









plaque formation and infectivity similar to wild type









loss of RNA replication, low VLP production with reversion



rescues the defect in I59N (compensatory), allows for virus

assembly/secretion

severely impaired RNA replication, low VLP production with reversion to I59Y



enhanced RNA replication, no effect on VLP production



loss of RNA replication, no effect on VLP production



enhanced RNA replication, no effect on VLP production



enhanced RNA replication, no effect on VLP production



enhanced RNA replication, no effect on VLP production



100% colony-forming efficiency, antigen staining less intense than WT

suggesting reduced WNR replication, weak expression of eGFP fluroescence

, infected Huh7 cells appeared healthy and dividing but were antigen positive,

produced smaller antigen positive foci than WT in vero cells, very little CPE

after 72hpi, not virulent in mice, IFN response delayed and deceased

compared to WT

100% colony-forming efficiency, antigen staining less intense than WT

suggesting reduced WNR replication, weak expression of eGFP fluroescence

,produced smaller antigen positive foci than WT in vero cells, very little CPE

after 72hpi, not virulent in mice

3.5% colony-forming efficiency, WT antigen staining efficiency, WT eGFP

fluroescence intensity, infected Huh7 cells appeared moribund like WT and

were antigen positive, produced somewhat smaller antigen postive foci than

WT but larger than other two NS2A mutations, CPE readily evident after

72hpi, virulent to mice and showed no reversion to WT









Page 58



Related docs
Other docs by linzhengnd
option strategy excel spreadsheet
Views: 3  |  Downloads: 0
Tips on Effective Listening
Views: 0  |  Downloads: 0
TO DOWNLOAD TEXT - Repairing The Breach
Views: 0  |  Downloads: 0
Power-Up Tested - Access Mobile
Views: 4  |  Downloads: 0
6502 Sell stone monuments and memorials
Views: 0  |  Downloads: 0
Sheet1 - Atlanta International School
Views: 2  |  Downloads: 0
AFRICAN UNION
Views: 0  |  Downloads: 0
By registering with docstoc.com you agree to our
privacy policy

You are almost ready to download!

You are almost ready to download!