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同步抑制棉籽 FAD2 1FatB基因表达的RNAi载体构建
引用本文:赵彦朋,李艳军,孙 杰,刘 峰.同步抑制棉籽 FAD2 1FatB基因表达的RNAi载体构建[J].西北农业学报,2013,22(10):54-61.
作者姓名:赵彦朋  李艳军  孙 杰  刘 峰
作者单位:(石河子大学 农学院,新疆石河子 832000)
基金项目:国家自然科学基金(31101094);兵团博士基金(2012BB004);石河子大学高层次人才项目(RCZX201006)。
摘    要:采用PCR技术,分别扩增获得靶标基因 FAD2 1FatB的功能区域片段426与501 bp,通过PCR引物引入特定的酶切位点,采用DNA标准重组技术与Gateway技术相结合的方法,以表达载体pBI121及RNA干扰载体pANDA35HK为基础,利用种子特异性启动子,成功构建出棉籽特异抑制双基因表达的RNAi载体。

关 键 词:FAD2  1  基因  FatB基因  Gateway技术  双基因沉默  载体构建

Construction of Plant Expression Vector for the Simultaneous Silencing of FAD2 1 and FatB Genes in Cotton Seeds
ZHAO Yanpeng,LI Yanjun,SUN Jie and LIU Feng.Construction of Plant Expression Vector for the Simultaneous Silencing of FAD2 1 and FatB Genes in Cotton Seeds[J].Acta Agriculturae Boreali-occidentalis Sinica,2013,22(10):54-61.
Authors:ZHAO Yanpeng  LI Yanjun  SUN Jie and LIU Feng
Abstract:To regulate fatty acid metabolism of cottonseed by simultaneous inhibition of FAD2 1 and FatB genes expression levels for improving the quality of cottonseed oil, the 426 bp and 501 bp fragments of Dual Target gene FAD2 1 and FatB were respectively amplified by PCR technology in this study firstly. Combining DNA recombinant technique with Gateway technology, the cottonseed specific Dual Target gene silencing RNAi vector was successfully constructed by introducing the seed specific promoter, based on the expression vector pBI121 and RNAi vector pANDA35HK. This study provided the foundation for further to apply for cotton genetic transformation and improve the quality of cottonseed fatty acids.
Keywords:FAD2 1  gene  FatB gene  Gateway technology  Dual Target gene silencing  Vector construction
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