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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

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【作者】 张伟尹峰何雯魏然刘春林阮颖

【Author】 Zhang Wei 1,2,Yin Feng 1,2,He Wen 1,2,Wei Ran 2,Liu Chunlin 2,Ruan Ying 1(1 College of Bioscience and Biotechnology,Hunan Agricultural University,Changsha 410128;2 Pre-state Key Laboratory for Germplasm Innovation and Resource Utilization of Crop,Changsha 410128)

【机构】 湖南农业大学生物科学技术学院作物种质创新和资源利用国家重点实验室培育基地

【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。

【Abstract】 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.

【基金】 国家自然基金面上项目(31071455)
  • 【文献出处】 中国农学通报 ,Chinese Agricultural Science Bulletin , 编辑部邮箱 ,2012年18期
  • 【分类号】S565.4
  • 【被引频次】6
  • 【下载频次】219
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