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.
出处
《作物杂志》
CAS
CSCD
北大核心
2014年第2期34-38,共5页
Crops
基金
转基因专项课题(2011ZX08011-003)
关键词
玉米
CRY1A
301基因
载体构建
原核表达
遗传转化
Maize
Cry1A. 301 gene
Vector construction
Prokaryotic expression
Genetic transformation
