【摘要】 目的:克隆并表达斜纹夜蛾核多角体病毒(Spodoptera litura multicapsid nucleopolyhe drovirus,SpltMNPV)ORF124基因的部分编码序列。方法:用PCR方法扩增目的基因序列片段,并将其克隆至原核表达载体pQE-30上,转化到Escherichia.coli M15[pREP-4]中进行诱导表达。结果:构建了pQE-tr124原核表达质粒,含有该质粒的大肠杆菌经IPTG诱导表达了一个与预期理论值相符的约为33kDa的蛋白。以Ni2+-NTA偶连抗体检测证明所表达的蛋白为带有组氨酸的融合蛋白。结论:成功表达了SpltMNPV ORF124的部分编码序列,该融合蛋白的成功表达为进一步深入研究基因的功能奠定了基础。更多还原

【Abstract】 Objective:To clone and express part of the code sequece of Spodoptera litura multicapsid nucleopolyhedrovirus ORF124 gene. Method:Target gene was amplified with PCR.The PCR product was cloned into the expression vector pQE-30. The resulting expression vector was transformed into Escherichia coli M15[pREP-4]. Result:Prokaryotic expression vector pQE-tr124 was constructed and a protein around 33kDa was expressed after induction with 1 mmol/L IPTG.Western blot analysis using Ni2+-NTA conjugate indicated that the protein contained 6×His tag. Conclusion: Truncated SpltMNPV ORF124 gene was epressed successfully.The expression of the fusion protein was suitable for further research of the gene function.更多还原