LMW-GS16基因表达载体的构建  

Construction of Plant LMW-GS16 Expression Vector

作　　者：武丽敏[1] 江千涛[1] 李琴[1] 魏育明[1] 郑有良[1] 

机构地区：[1]四川农业大学小麦研究所

出　　处：《吉林农业大学学报》2005年第4期381-384,共4页Journal of Jilin Agricultural University

基　　金：国家"863"计划项目(2003AA207100);长江学者和创新团队发展计划项目(2003AA207100)

摘　　要：采用PCR方法对来自大麦的LMW-GS16基因进行修饰,在基因5′及3′端分别加入构建表达载体所需的BamHⅠ和SacⅠ限制酶切位点,并将其连接在相应酶切的质粒pBI121上,构建成LMW-GS植物表达载体pBI121-16。重组质粒转化到E.coliDH5α和农杆菌LBA4404中,通过Kanamycin筛选阳性克隆,用PCR方法进行鉴定。对照组转化率为零,转化组阳性克隆特异地扩增出900 bp片段,与目标基因DNA大小一致,转化率为1.0×106/μg DNA。LMW - GS16 gene from barley（ Hordeum vulgare L.） has been modified by PCR method. Through primers design, a site BamH I and a Sac I site have been added to 5＇-end and 3＇-end of LMW - GS16 gene respectively. PCR production of LMWGS16 Was digested with restriction endonucleases, and the acquired LMW - GS16 was ligated to engineered plasmid pBI121 which was digested with appropriated restriction eddonucleases, and therefore a plant expression vector for pBI121 - 16 was constructed. The recombined plasmid was transformed into DH5α and LBA4404 respectively. PCR production of resistant kanamycin clones was 900 bp which was the same as the anticipation. The transform frequency was zero in control and 1.0 x 10^6/μg DNA in transformed group.

关 键 词：表达载体 LMW-GS基因 pBI121-16 

分 类 号：Q782[生物学—分子生物学]