杆状病毒SpltMNPV ORF124基因截短片段的克隆与表达  

Cloning and Expression of Truncated Spodoptera litura Multicapsid Nucleopolyhedrovirus ORF124 Gene

作　　者：李玲玲[1,2] 庞义[1] 

机构地区：[1]中山大学有害生物控制与资源利用国家重点实验室,广东广州510275 [2]华南师范大学激光生命科学教育部重点实验室,广东广州510631

出　　处：《生物技术》2009年第2期3-5,共3页Biotechnology

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。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 Ni^2＋-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.

关 键 词：斜纹夜蛾核多角体病毒 ORF124 克隆 原核表达 

分 类 号：Q939.96[生物学—微生物学] S182[农业科学—农业基础科学]