斜纹夜蛾核多角体病毒VP39-GST融合蛋白原核表达

Expression of Spodoptera litura Multicapsid Nucleopolyhedrovirus VP39-GST Fusion Protein in Escherichia coli

用PCR的方法扩增得到斜纹夜蛾核多角体病毒(Spodopteralituramulticapsidnucleopolyhedrovirus,SpltMNPV)vp39全长基因。将其克隆至原核表达载体pGEX-4T-1上,构建重组表达质粒pGEX-4T-vp39,转化Escherichia.coliBL21(DE3),在1mmol/L异丙基硫代-β-D-半乳糖苷(IPTG)诱导下超量表达了与理论预测值相符的一个约60kDa的VP39-GST融合蛋白。VP39-GST融合蛋白的成功表达为进一步研究VP39在病毒侵染过程中与宿主细胞成分或病毒粒子蛋白间的相互作用奠定了基础。

The ORF of Spodoptera litura multicapsid nucleopolyhedrovirus SpltMNPV vp39 gene was obtained by PCR method from SpltMNPV genomic DNA. The PCR product was cloned into pMD18-T vector to get the recombinant plasmid （pMD18-vp39）. The vp39 gene was recombined in vitro with prokaryotic expression vector pGEX-4T-1 and transformed into Escherichia colt BL21 （ DE3 ）, The BL21 （ DE3 ） strain, containing vp39 recombinant plasmid, expressed a 60kDa fusion protein after the induction with 1 mmol/L IPTG （ Isoprophylthio- β-D-galacloside ）. The expression of the VP39-GST fusion protein was suitable for further analysis of the interaction between baculovirus and its host.