To improve the expression of the foreign genes in recombinant goat poxvirus (GPV) and screen for strong promoter,the recombinant plasmids were constructed with the firefly luciferase (Fluc) gene as report gene under control of 8 poxvirus promoters,respectively,which included the promoters of Amsacta moorei entomopoxvirus 42K (42K),poxvirus synthetic late 441 (SL441),poxvirus synthetic late 454 (SL454),vaccine virus (VV) early/late P7.5 (P7.5),VV late P11 (P11),VV H6 (H6),VV synthetic early/late (E/L) and synthetic late-early optimized (LEO).The recombinant plasmids were transfected into four animal cells [DF-1 (chicken),LT (sheep),BHK-21 (hamster) and Vero (monkey)] pre-infected with GPV for 1 hour,respectively,and plasmid pRL-TK expressing Renilla luciferase (Rluc) was used as internal control.Luciferase activities were then detected at 24 hours.The results showed that in LT cells,SL441 promoter was the strongest;in DF-1 cells,LEO promoter was the strongest,and was 1.19 to 14.46 times to others;in Vero cells,the transcription efficiency of H6 and SL441 promoters were similar,and were 1.21 to 11.24 times to others;in BHK-21 cells,H6 promoter was significantly stronger than the others.Taken together,we identified different promoters' activities in four cells infected by GPV,which established the foundation for improving immune efficacy of GPV recombinant vaccine.
NETL
NSTL
维普
万方数据
