红笛鲷SR-BⅠ基因的原核表达及条件优化  

Prokaryotic Expression and Condition Optimization of SR-BⅠ Gene in Lutjanus sanguineus

作　　者：王一帆 陈子茵 贾新蕾 黄增朝 吕静 黄郁葱[1] WANG Yifan(Fisheries College of Guangdong Ocean University/Provincial Key Laboratory of Pathogenic Biology and Epidemiolo-gy for Aquatic Economic Animals of Guangdong&Key Laboratory of Control for Diseases of Aquatic Economic Ani-mals of Guangdong Higher Education Institute,Zhanjiang 524088,China)

机构地区：[1]广东海洋大学水产学院/广东省水产经济动物病原生物学及流行病学重点实验室暨水产经济动物病害控制广东省普通高等学校重点实验室,广东湛江524088

出　　处：《安徽农学通报》2021年第3期1-5,48,共6页Anhui Agricultural Science Bulletin

基　　金：广东省自然科学基金项目(2016A030313748);大学生创新创业训练计划(CXXL2019139);南方海洋科学与工程广东实验室(湛江)自主项目(ZJW-2019-06)。

摘　　要：根据GenBank上登录的红笛鲷SR-BⅠ基因设计引物,采用RT-PCR方法扩增该基因胞外段序列,然后将该基因胞外段序列定向克隆到原核表达载体pET-28a(+)中,将重组质粒转入大肠杆菌BL21进行IPTG诱导表达,并对重组表达菌株的表达条件进行优化。结果显示,成功构建了原核表达载体pET28a-SR-BI,并能在大肠杆菌BL21中表达,表达的融合蛋白分子量为50.0ku。融合蛋白的最佳诱导表达温度、IPTG浓度和时间分别为16℃、0.05mmol/L和8h。研究结果为进一步研究SR-BI的功能奠定了基础。A pair of primers were designed based on Scavenger receptor class B type Ⅰ(SR-BⅠ)gene sequence published in GenBank. The extracellular region sequence of SR-BI gene was amplified by RT-PCR and then inserted into the pET-28 a(+)vector to construct prokaryotic expression plasmid. The recombinant plasmid was transformed into E.coli BL21 to overexpressed in the presence of isopropyl-β-D-thiogalato pyranos ide(IPTG), and the expression conditions of the recombinant strain were optimized. The results showed that the prokaryotic expression plasmid pET28 a-SR-BI was successfully constructed and expressed in E. coli BL21. The molecular weight of the expressed recombinant fusion protein was 50.0 ku. The optimal induction expression temperature, IPTG concentration and induction time of the recombinant fusion protein was 16℃, 0.05 mmol/L and 8 h, respectively. The research results laid the foundation for the further research on the functions of SR-BI.

关 键 词：红笛鲷 SR-BⅠ基因 原核表达 条件优化 

分 类 号：S942.1[农业科学—水产养殖]