文献类型：期刊文献

中文题名：转人Cu,Zn-SOD基因马铃薯的研究

英文题名：Expression of the hCu,Zn-SOD Gene in Potato

作者：刘晓鹏[1,2];姜宁[2];田阼茂[2];吕胜安[2];袁勤生[1]

机构：[1]华东理工大学生物反应器工程国家重点实验室,生物化学研究所,上海200237;[2]中国南方马铃薯中心,湖北445000

年份：2003

卷号：29

期号：3

起止页码：255

中文期刊名：华东理工大学学报（自然科学版）

外文期刊名：Journal of East China University of Science and Technology

收录：CSTPCD;;EI(收录号：2003307568883);Scopus;北大核心:【北大核心2000】；CSCD:【CSCD2011_2012】；

语种：中文

中文关键词：SOD;hCu,Zn—SOD;农杆菌;基因克隆

外文关键词：SOD; hCu, Zn SOD; Agrobacterium tumefaciens; gene cloning

摘要：采用 RT-PCR技术从人肝总 RNA中分离扩增了 490 bp的人铜锌超氧化物歧化酶(h Cu,Zn-SOD)基因的 c DNA序列 ,进行了序列测定 ,结果和文献报道的一致 ;构建了 Ca Mv3 5 S启动子驱动 h Cu,Zn-SOD基因的植物表达载体 PBICu SOD;用三亲交配法将重组质粒 PBICu SOD转化到农杆菌 LBA440 4,转化马铃薯得到转基因植株。PCR和 Southern-blot分析证明 ,h Cu,Zn-SOD基因已整合到马铃薯基因组中。Western blot分析表明 ,h Cu,Zn-SOD基因在转基因马铃薯植株中得到表达。用 NBT光还原法测定转基因马铃薯植株中的 SOD酶活力 ,叶的总酶活为1 0 66.6U/ g(湿重 ,下同 ) ,块茎的总酶活为 1 0 1 1 U/ g。
The cDNA encoding human Cu/Zn superoxide dismutase was amplified by RT PCR using the total RNA of human liver as the template, and was cloned into the plasmid pBluescript SK. The sequence of the cloned h SOD cDNA was consistent with the reported data. The h SOD cDNA was cloned into plant expression vector PBICuS. The recombinant plasmid PBICuS was introduced into Agrobacterium tumefaciens LBA4404. Potato plants were transformed via Agrobacterium tumefaciens mediated procedure. It was confirmed by PCR and Southern blot analysis that the human Cu, Zn SOD gene was expressed in potato at high level. The expression product was characterized by Western blot analysis. The SOD activity of transgenic potatoes was detected by the inhibition of nitro tetrazolium (NBT) reduction. The activity in tubes was 1 011 U/g(FW) and the activity in leaves was 1 066.6 U/g(FW).