苏云金芽孢杆菌Cry2Aa蛋白的体外表达及纯化  

In Vitro Expression and Purification of Bacillus thuringiensis Cry2Aa Protein

作　　者：高洁荣[1,2] 何颖[2] 邹泽红[2] 艾云灿[1] 

机构地区：[1]中山大学生命科学院/有害生物控制与资源利用国家重点实验室,广州510275 [2]广州医学院第二附属医院/呼吸疾病国家重点实验室变态反应研究室/广州市过敏反应与临床免疫重点实验室,广州510260

出　　处：《湖北农业科学》2013年第3期702-705,共4页Hubei Agricultural Sciences

基　　金：国家自然科学基金(30771240);国家科技重大专项重大课题(2011ZX08011-005);国家科技重大专项重点课题(2009ZX08011-004B);呼吸疾病国家重点实验室(广州医学院)课题研究基金

摘　　要：根据大肠杆菌密码子偏好性对苏云金芽孢杆菌(Bacillus thuringiensis)cry2Aa基因进行优化,并合成全长基因,将cry2Aa基因克隆到表达载体pET-44a上,转化到大肠杆菌Rosetta中进行诱导表达,优化表达条件,采用亲和层析和SDS-PAGE胶回收纯化,Western blot鉴定回收产物。结果表明,IPTG浓度为0.50 mmol/L、27℃诱导4 h时Cry2Aa蛋白表达量最高,SDS-PAGE胶回收Cry2Aa纯化蛋白所得的产量高于亲和层析法。cry2Aa gene of Bacillus thuringiensis was optimized according to codon preference of Escherichia coli,which was then synthesized and cloned into expression vector pET44a and transformed into E.coli Rosetta strain.The expression of Cry2Aa protein was induced under optimized conditions.The expressed protein was purified by affinity chromatography and SDS-PAGE gel extraction,and identified by Western blot.The results showed that expression of Cry2Aa protein was the highest when induced by 0.50 mmol/L IPTG at 27 ℃ for 4 h.Yield of purified recombinant protein Cry2Aa obtained by SDSPAGE gel extraction was higher than that of affinity chromatography.

关 键 词：苏云金芽孢杆菌(Bacillus thuringiensis) Cry2Aa蛋白 表达 纯化 

分 类 号：S[农业科学]