甘蓝型油菜iMyAP9、iMyAP12基因过表达载体的构建及转化拟南芥  

The Over-expression Vector Construction of BnaiMyAP9 and BnaiMyAP12 and Transformation of Arabidopsis thaliana

作　　者：张伟[1,2] 尹峰[1,2] 何雯[1,2] 魏然 刘春林 阮颖[1] 

机构地区：[1]湖南农业大学生物科学技术学院,长沙410128 [2]作物种质创新和资源利用国家重点实验室培育基地,长沙410128

出　　处：《中国农学通报》2012年第18期174-178,共5页Chinese Agricultural Science Bulletin

基　　金：国家自然基金面上项目(31071455)

摘　　要：诱导型黑芥子酶协助蛋白(iMyAP)往往与黑芥子酶和黑芥子酶结合蛋白一起以复合体的形式存在,对植物的防御系统有着重要的作用。为了研究黑芥子酶协助蛋白在植物响应非生物胁迫方面的功能,通过以pBI121为载体,分别构建了CaMV35S::iMyAP9和CaMV35S::iMyAP12,浸花法转化拟南芥,用卡那霉素对转化植株进行筛选,并对抗性苗进行PCR检测,初步得到67株转基因植株;提取转基因拟南芥中RNA进行RT-PCR表达分析。结果表明,iMyAP9基因和iMyAP12基因在随机挑选的转基因拟南芥中获得了有效的表达。拟南芥转基因植株的获得为进一步探讨iMyAP9基因、iMyAP12基因的生物学功能奠定了基础。Inducible Myrosinase-associated protein（iMyAP） usually occur in complexes with Myrosinases and Myrosinases-binding protein plays important roles in the plant ’ s defense system.To study the functions of iMyAP in abiotic stress responses,we constructed two plant over-expression vectors CaMV35S：：iMyAP9 and CaMV35S：：iMyAP12.Genetic transformation of Arabidopsis was performed by floral dip method.The T 0 seeds were screened on medium with kanamycin and the anti-kanamycin plants were tested by PCR to confirm if anti-Kan plants were transgenic plants.67 transgenic plants were obtained.The results of RT-PCR showed that iMyAP9 and iMyAP12 genes efficiently expressed in the transgenic arabidopsis plants,which the results lay a foundation for further studying the biological function of iMyAP9 and iMyAP12 genes.

关 键 词：油菜 黑芥子酶协助蛋白 iMyAP基因 转基因拟南芥 

分 类 号：Q7[生物学—分子生物学]