关键词: 兔轮状病毒, VP7基因, 杆状病毒表达

Abstract: Objective To express the LaRV VP7 protein gene with Bac-to-Bac baculovirus expression system. Method According to LaRV VP7 gene sequence in Genbank, a pair of specific primers was designed and synthesized. The VP7 ORF gene was amplified by RT-PCR., and then inserted into the transfer vehicle pFastbacHTa to obtain the recombinant transfer vectors pFastBHa-VP7, which was used to transform into E. coli DHlOBac, producing specific transposition with Bacmid to yield the recombinant shuttle vehicles, Bacmid-VP7. Then the Bacmid-VP7 was further used to transfect Sf9 insect cells by lipofectin to generate recombinant baculovirus expressing VP7 recombination protein. Results The recombinant baculovirus and Bacmid-VP7 was obtained. The IFA and Western blotting detection confirmed that the gene had been expressed in sf9 cells, and the expression product could specially react to the sera of LaRV. Conclusion The LaRV VP7protein is expressed successfully by Bac-to-Bac baeulovirus expression system, which may lay foundation for development of diagnostic technique.

Key words: Lapine Rotavirus, VP7 gene, Baculovirus expression