贵州农业科学2025，Vol.53Issue(4)：66-72,7.DOI:10.3969/j.issn.1001-3601.2025.04.008
家蝇丝氨酸蛋白酶抑制剂Serpin2的诱导表达条件优化及原核重组蛋白纯化
Optimization of Induced Expression Conditions of Serpin2 in Musca domestica and Purification of Prokaryotic Recombinant Protein
摘要
Abstract
[Objective]A large number of purified recombinant proteins of Serpin2,a serine protease inhibitor in Musca domestica were obtained by induced expression to lay the foundation for the subsequent in-depth study of the immunomodulatory function of Serpin in M.domestica,a model insect.[Method]The prokaryotic expression system of Serpin2 in M.domestica was constructed using pET-28a(+)as the expression vector and introduced into Escherichia coli(BL21),and the optimal induction conditions were explored by setting different IPTG concentrations,different induction temperatures and different induction times,under which a large number of prokaryotic recombinant proteins of Serpin2 were induced and purified,and the purified recombinant Serpin2 protein was identified by SDS-PAGE gel electrophoresis and protein spectroscopy.[Result]The recombinant plasmid of pET28a(+)-Serpin2 was successfully constructed and transformed into competent cells.It was confirmed that the recombinant protein of Serpin2 in M.domestica was optimally expressed in E.coli BL21 at a final concentration of 0.2 mmol/L IPTG for 15 h at 30℃,and both in supernatant and inclusion body.The purified protein was identified as Serpin2 recombinant protein of M.domestica.[Conclusion]A large number of purified Serpin2 recombinant proteins of M.domestica are obtained under the optimum induction conditions(30℃,0.2 mmol/L IPTG and 15 h).关键词
家蝇/丝氨酸蛋白酶抑制剂2/原核表达优化/蛋白纯化/重组质粒/重组蛋白Key words
Musca domestica/Serpin2/prokaryotic expression optimization/protein purification/recombinant plasmid/recombinant protein分类
农业科技引用本文复制引用
王玮,王丽,国果,修江帆,尚小丽,张昌容..家蝇丝氨酸蛋白酶抑制剂Serpin2的诱导表达条件优化及原核重组蛋白纯化[J].贵州农业科学,2025,53(4):66-72,7.基金项目
2022年贵州省大学生创新创业训练计划项目(S202210660058) （S202210660058）
国家重点研发计划项目"重大病虫害防控综合技术研发与示范"专项"天敌昆虫资源发掘利用与新产品创制"(2023YFD1400603) （2023YFD1400603）
贵州省科技计划项目(黔科合基础-ZK[2022]一般 377) （黔科合基础-ZK[2022]一般 377）
国家重点研发计划"政府间国际科技创新合作"重点专项"中欧农作物病虫害综合治理关键技术创新合作研究"(2023YFE0104800) （2023YFE0104800）
