Gateway技术构建棉花GhSAMDC基因植物表达载体  

Construction of Plant Expression Vector for GhSAMDCGene of Cotton Using Gateway Technology

作　　者：耿卫东[1] 李艳军[1] 张新宇[1] 孙杰[1] 朱华国[1] 

机构地区：[1]石河子大学农学院/新疆兵团绿洲生态农业重点实验室,石河子832003

出　　处：《石河子大学学报（自然科学版）》2012年第2期138-141,共4页Journal of Shihezi University(Natural Science)

基　　金：新疆兵团博士资金项目(2011BB004);石河子大学高层次人才项目(RCZX200905)

摘　　要：为了进一步转化植物,研究棉花GhSAMDC基因在植物抗逆过程中所起作用,用Gateway技术构建棉花Gh-SAMDC基因RNA干扰载体和超表达载体。通过BP反应将干扰片段重组到植物表达载体Hellsgate4中,分别用XbaⅠ和XhoⅠ单酶切,酶切片段大小一致,表明GhSAMDC基因RNA干扰载体构建成功。通过BP反应和LR反应将超表达目的片段重组到植物表达载体pGWB17中,经HindⅢ和SacⅠ双酶切验证,酶切片段大小与目的片段一致,表明GhSAMDC基因超表达载体构建成功。As one of the key enzymes in polyamine biosynthesis,S-adenosylmethionine Decarboxylase （SAMDC） has an effect on response environments stress of plants by accommodating the content of polyamines. We designed primers according to the cD- NA sequence of GhSAMDC gene. Through polymerase chain reaction,we obtained 237 bp fragment for RNAi vector construction and 1159 bp fragment for over-expression vector construction. The 237 bp fragment was recombined with Hellsgate4 vector by BP reaction. After Xba Ⅰ and Xho Ⅰ restriction enzyme digestion,we got two fragments with same size,which suggested that the RNAi vector was constructed successfully. In the same way, the 1159 bp fragment was recombined with the pGWB17 vector by BP reaction and LR reaction. Digestion by HindⅢ and Sac Ⅰ restriction enzyme indicated that the over-expression vector Gh- SAMDC-pGWB17 was constructed successfully. The constructed vectors were transformed into agrobacterium,which would lay a foundation for further study of GhSAMDC function.

关 键 词：GATEWAY技术 SAMDC 植物表达载体 

分 类 号：S336[农业科学—作物遗传育种] S562[农业科学—农艺学]