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Cloning and Expression of a Baculovirus J Domain Protein
Gene in E. coli and Preparation of Antiserum |
| Wang Lihua Yu Jianxiu Yin Chong Li Zhaofei Pang Yi** |
| (State Key Laboratory for Biocontrol and Institute of Entomology, Zhongshan University, Guangzhou 510275, China) |
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Abstract Abstract: Spodoptera litura multicapsid nucleopolyhedrovirus bjdp (baculovirus J domain protein ) gene is the only gene encoding a J domain protein homology in known baculovirus genome. Sequence analysis indicated that the N terminus of the gene production had a conserved J domain. With the specific primers which designed on bjdp gene reported recently, the coding region was amplified from SpltMNPV genome DNA via PCR . The PCR product was cloned into the pGEM-T Easy vector to get the recombinant plasmid (pGT-B). The bjdp gene was recombined in vitro with expression vector pQE30 and transformed into E.coli M15[pREP4]. The M15[pREP4] containing bjdp recombinant plasmid expressed a 36 kD 6×His-tag fusion protein after the induction with 1mmol/L IPTG. The fusion protein was purified with Ni-NTA resin column and used as the immunogen to raise a BJDP-specific antiserum. Western blot analysis indicated that the antiserum could react with the corresponding protein existed in SpltMNPV-infected S1-zsu-1 cells.
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Received: 01 January 1900
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