Substitution of the premembrane and envelope protein genes of Modoc virus with the homologous sequences of West Nile virus generates a chimeric virus that replicates in vertebrate but not mosquito cells

Table 1 PCR products generated during the construction of full-length flavivirus chimeric DNAs

Reaction no.	Reaction type	Primers (forward, reverse)	PCR product
Reaction no.	Reaction type	Primers (forward, reverse)	Name	Size (bp)
1a	PCR	M-F1, MW-R1	MW1’	191
1b		M-F1, MC-R1	MC1’	194
1c		M-F1, MWi-R1	MW1	523
1d		M-F1, MCi-R1	MC1	521
2a	RT-PCR	MW-F2, MW-R2	MW2’	2,415
2b		MC-F2, MC-R2	MC2’	2,167
2c		MWi-F2, MWR2	MW2	2,066
2d		MCi-F2, MCR2	MC2	1,777
3a,c	PCR	MW-F3, M-R3	MW3	2,575
3b,d	PCR	MC-F3, M-R3	MC3	2,580
4a-d	PCR	M-F4, M-R10600	M4	6,227
5a	Fusion-PCR	M-F1, MW-R2	MW5’	2,563
5b		M-F1, MC-R2	MC5’	2,320
5c		M-F1, MW-R2	MW5	2,542
5d		M-F1, MC-R2	MC5	2,251
6a	Fusion-PCR	M-F1, M-R3	MW6’	5,100
6b		M-F1, M-R3	MC6’	4,854
6c		M-F1, M-R3	MW6	5,079
6d		M-F1, M-R3	MC6	4,785
7a	Fusion-PCR	T7MOD-F, M-R10600	fpMODV-WNV(C-prM-E)	10,730
7b		T7MOD-F, M-R10600	fpMODV-WNV(prM-E)	10,708
7c		T7MOD-F, M-R10600	fpMODV-CxFV(C-prM-E)	10,484
7d		T7MOD-F, M-R10600	fpMODV-CxFV(prM-E)	10,415

Reactions ending with a, b, c and d were used to generate fpMODV-WNV(C-prM-E), fpMODV-CxFV(C-prM-E), fpMODV-WNV(prM-E) and fpMODV-CxFV(prM-E), respectively.

ISSN: 1743-422X