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Mi基因过表达载体和16D10基因RNAi载体的构建及转化

Construction of Mi Gene Over-Expression Vector and 16D10 Gene RNAi Vector and Their Transformation

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【作者】 吴倩倩胡敏伦钟云闫化学姜波吴波钟广炎高峰

【Author】 WU Qianqian;HU Minlun;ZHONG Yun;YAN Huaxue;JIANG Bo;WU Bo;ZHONG Guangyan;GAO Feng;Guangdong Provincial Key Laboratory of Biotechnology for Plant Development∥School of Life Sciences,South China Normal University;Institute of Fruit Tree Research,Guangdong Academy of Agricultural Sciences;Key Laboratory of South Subtropical Fruit Biology and Genetice Resource Utilization∥Guangdong Province Key Laboratory of Tropical and Subtropical Fruit Tree Research;

【通讯作者】 钟云;高峰;

【机构】 广东省植物发育生物工程重点实验室∥华南师范大学生命科学学院广东省农业科学院果树研究所农业部南亚热带果树生物学与遗传资源利用重点实验室∥广东省热带亚热带果树研究重点实验室

【摘要】 文中采用PCR技术从番茄中克隆获得了Mi基因,采用反转录PCR(RT-PCR)技术从来源于红橘(Citus reticulate)的根结线虫中克隆获得了16D10基因,采用DNA重组技术构建了Mi基因的过表达载体和16D10基因的RNAi载体,分别命名为pB-Mi和pB-16DR.通过根瘤农杆菌介导的遗传转化,获得了经PCR检测为阳性的沙田柚转基因植株.这一结果将为研究Mi基因过表达和16D10基因的RNA干扰对柑橘根结线虫病的抗性影响提供试材和技术支持.

【Abstract】 Mi gene was cloned from tomato with PCR amplification and 16 D10 gene was cloned from root knot nematode in Citus reticulate with reverse-transcription PCR(RT-PCR) amplification. The Mi gene over-expression vector(named pB-Mi) and 16 D10 gene RNAi vector(named pB-16 DR) were constructed with DNA molecular recombination technology. Putative transgenic plantlets,which were confirmed by PCR analysis,were obtained through genetic transformation mediated by Agrobacterium tumefaciens. The results can provide material and technical basis for exploring the effect of the over-expression of Mi gene and RNA interference of 16 D10 gene on the resistance of Citrus root knot nematode disease.

【基金】 国家自然科学基金项目(31572113);广东省科技计划项目(2014B020202009,2014B020203003,2016B020201006,2017A070702005)
  • 【文献出处】 华南师范大学学报(自然科学版) ,Journal of South China Normal University(Natural Science Edition) , 编辑部邮箱 ,2018年04期
  • 【分类号】S436.66
  • 【下载频次】285
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