水稻抗病相关基因Os01g0620100的克隆及表达载体构建
首发时间:2014-04-25
摘要:根据拟南芥抗病相关基因She1序列,获得水稻一个同源基因Os01g0620100,以水稻品种日本晴总RNA为模板,反转录得到cDNA,根据水稻Os01g0620100基因序列设计1对特异性引物进行PCR扩增,回收PCR产物连接到pEASY-T1 Simple载体中,筛选阳性克隆进行测序,然后将基因片段亚克隆到pCAMBIA1301a载体中,构建了过表达载体pCAMOs01g0620100。同时扩增了该基因的一段400bp左右片段,通过正反向连接,成功构建了干涉载体,这两个载体的构建对于研究水稻Os01g0620100基因的功能具有重要的意义。
关键词: 水稻 Os01g0620100基因 过表达载体 干涉载体 构建
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Cloning of Os01g0620100 Gene and Vectors Construction in Rice
Abstract:With the total RNA of japonia rice variety Nipponbare as templates,reversing transcription for cDNA,a pair of specific premiers was designed according to cDNA sequence of Os01g0620100 gene in rce for amplification.The reclaimed PCR products were connected into pEASY-T1 simple vector and the cloning pEASY-TIOs01g0620100 of Os01g0620100 gene in rice was obtained through screening.The cloning fragment and pCAMBIA1301a vector was connected and successfully constructed the overexpression vector pCAMOs01g0620100.Then,a interference fragment was made the amplication,and the interference vector pCAMAOs01g0620100 was constructed.It is of the vital significance of the two vectors for researching the function of Os01g0620100 gene in rice.
Keywords: Rice Os01g0620100 gene Overexpression vector Interference vector Construction
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