抗玉米螟基因Cry1A.301原核表达和玉米遗传转化  

Prokaryotic Expression and Genetic Transformation of Corn Borer Resistance Gene Cry1A. 301 in Maize

作　　者：王慧[1,2] 杨小艳[3] 翁建峰[2] 宋新元[4] 王大铭[4] 王振华[1] 李新海[2] 

机构地区：[1]东北农业大学农学院,黑龙江哈尔滨150030 [2]中国农业科学院作物科学研究所/作物分子育种国家工程实验室,北京100081 [3]重庆市农业科学院,重庆401329 [4]吉林省农业科学院,吉林长春130124

出　　处：《作物杂志》2014年第2期34-38,共5页Crops

基　　金：转基因专项课题(2011ZX08011-003)

摘　　要：增强玉米品种的抗虫性是减少玉米螟为害、提高玉米产量的有效途径。通过密码子优化改造获得抗虫基因Cry1A.301,构建原核表达载体pET30a-Cry1A.301,进行蛋白免疫学、ELISA检测以及玉米螟抗性生测。结果表明,原核表达载体中Cry1A.301蛋白高效表达,且7d后杀虫效率达100%。通过Cry1A.301基因与植物表达载体CPB连接,构建CPB-35S∶∶Cry1A.301-35S∶∶bar载体进行农杆菌介导的玉米萌动胚遗传转化,得到12株T0代转基因植株。T0代植株叶片中目的基因、筛选标记bar基因的PCR检测与蛋白免疫学检测结果表明,Cry1A.301基因已经整合到玉米基因组并表达。Development and application of resistant maize varieties can reduce the damage caused by corn bor- er effectively. In this study, a new insect-resistant gene Cry1A. 301 was obtained by optimizing the codon. Pro- karyotic expression vector pET30a-Cry1A. 301 was constructed. The results of immuno strip test, ELISA test, and protein detection showed that there was a high expression level of Cry1A. 301 protein. Meanwhile, seven days after feeding corn borers with Cry1A. 301 protein the insecticidal rate reached 100%. Using plant express vector CPB-35S∷Cry1A. 301-35S∷bar, twelve transgenic plants in TO generation were obtained through Agrobacterium mediated genetic transformation in maize germinating embryo. PCR assay for gene Cry1A. 301 and bar in the transgenic plant leaves and immuno strip test suggested that gene Cry1A. 301 had been integrat- ed into the maize genome, and protein was expressed.

关 键 词：玉米 CRY1A 301基因 载体构建 原核表达 遗传转化 

分 类 号：S513[农业科学—作物学]