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

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.