核桃JrLFY基因表达载体的构建及工程菌的筛选  

Construction of Expression Vector and Selection of Engineering Strains from JrLFY Gene of Juglans regia L.

作　　者：何富强[1] 王红霞[2] 张志华[2] 崔彬彬[1] 

机构地区：[1]保定学院生化系,河北保定071000 [2]河北农业大学山研所,河北保定071001

出　　处：《湖北农业科学》2017年第8期1573-1576,共4页Hubei Agricultural Sciences

基　　金：保定市科学技术研究与发展指导计划项目(13ZN018);保定学院博士基金项目(2011Z01);国家自然科学基金项目(31300562);河北省自然科学基金面上项目(C2013104053);保定学院首批科研团队项目(KYTD2013001)

摘　　要：为了研究核桃JrLFY基因的功能,利用RT-PCR技术扩增JrLFY基因ORF,设计含NdeⅠ/KpnⅠ酶切位点的引物获得该基因CDS区片段,用限制性内切酶NdeⅠ/KpnⅠ双酶切p RI 101-AN表达载体,利用In-Fusion技术成功构建p RI 101-AN·JrLFY表达载体,并将该表达载体转化农杆菌GV3101感受态细胞,成功获得工程菌,为其基因缺失或过表达等试验提供技术支持。To make clear the function of JrLFY gene, its open reading frame （ORF） was cloned by RT-PCR in this paper. Then the CDS fragment was amplified by primers containing specific enzyme sites. The purified PCR product and the plant expression vector pRI 101-AN was digested by the restricted enzyme Nde I/Kpn [ . The final expression vector pRI 101- AN.JrLFY was constructed successfully by In-Fusion technique and transferred into agrobacterium stain GV3101. The genetic engineering strains were selected successfully, which will provide the technical support for the experiment such as gene dele- tion and gene overexpression and so on.

关 键 词：核桃 JrLFY基因 表达载体 工程菌 

分 类 号：S664.1[农业科学—果树学] Q78[农业科学—园艺学]