同步抑制棉籽FAD21-与FatB基因表达的RNAi载体构建  

Construction of Plant Expression Vector for the Simultaneous Silencing of FAD21- and FatBGenes in Cotton Seeds

作　　者：赵彦朋[1] 李艳军[1] 孙杰[1] 刘峰[1] 

机构地区：[1]石河子大学农学院,新疆石河子832000

出　　处：《西北农业学报》2013年第10期54-61,共8页Acta Agriculturae Boreali-occidentalis Sinica

基　　金：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FAD21-与FatB的功能区域片段426与501bp,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。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 frag- ments 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 spe- cific Dual-Target gene silencing RNAi vector was successfully constructed by introducing the seed spe cific 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.

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

分 类 号：S562[农业科学—作物学]