Expression of envelope protein of Zika virus in baculovirus expression system / 中华实验和临床病毒学杂志

Objective@#To express envelope protein of ZIKA virus in baculovirus expression system.@*Methods@#Full-length E gene of ZIKA virus was obtained by DNA synthesis and inserted into vector pFastBac1. The constructed recombinant baculovirus transfer vector pFB1-E was transformed to competent DH10Bac cells. The obtained skeleton plasmid rBacmid-E was transfected to sf9 cells, and the constructed recombinant baculovirus rBac-E was determined for titer, for insertion of E gene by PCR, and for expression of E protein by IFA and Western blotting.@*Results@#PCR proved that skeleton plasmid rBacmid-E was constructed correctly. The titer of rBac-E of passage 3 was 2.58×105pfu/ml. The genome of infected cells virus was extracted, the gene band at length of 3 830 bp was observed after PCR amplification. Indirect immunofluorescence of the infected cells showed the specific green fluorescence, 55×103specific band was determined by Western blotting identification in the cell pellet of the infected recombinant baculovirus rBac-E.@*Conclusions@#The recombinant baculovirus with E gene of ZIKA virus was successfully constructed, which laid a foundation of further study on the function of E protein and the vaccine of ZIKA virus.

The expression of hepatitis C virus truncated core and EGFP fusion gene in Sf9 cells / 临床检验杂志

Objective To construct recombinant baculovirus expression vector containing hepatitis C virus(HCV) truncated core gene and EGFP gene,and to study the antigenicity of the fusion protein expressed in Sf 9 cells.Methods PCR-amplified HCV truncated core gene and EGFP gene were cloned into the transposed vector pFastBac1 to construct a recombinant plasmid pFastCt-EGFP,by which E.coli DH10Bac was transformed to get the recombinant BacmidCt-EGFP.Insect Sf 9 cells were transfected with BacmidCt-EGFP and the expression of fusion protein Ct-EGFP was screened by SDS-PAGE and Western blotting.Results SDS-PAGE and Western blot analysis showed that fusion protein Ct-EGFP was expressed with the molecular mass of 40 kD.ELISA results showed that the fusion protein reacted with 15 of 28(54%) anti-HCV positive sera.Conclusion The fusion protein Ct-EGFP was expressed in insect Sf9 cells and showed partial antigenicity.