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    刘晓鹏, 姜宁, 田阼茂, 吕胜安, 袁勤生. 转人Cu,Zn-SOD基因马铃薯的研究[J]. 华东理工大学学报(自然科学版), 2003, (3): 255-258.
    引用本文: 刘晓鹏, 姜宁, 田阼茂, 吕胜安, 袁勤生. 转人Cu,Zn-SOD基因马铃薯的研究[J]. 华东理工大学学报(自然科学版), 2003, (3): 255-258.
    Expression of the hCu,Zn-SOD Gene in Potato[J]. Journal of East China University of Science and Technology, 2003, (3): 255-258.
    Citation: Expression of the hCu,Zn-SOD Gene in Potato[J]. Journal of East China University of Science and Technology, 2003, (3): 255-258.

    转人Cu,Zn-SOD基因马铃薯的研究

    Expression of the hCu,Zn-SOD Gene in Potato

    • 摘要: 采用RT—PCR技术从人肝总RNA中分离扩增了490bp的人铜锌超氧化物歧化酶(hCM,Zn—SOD)基因的cDNA序列,进行了序列测定,结果和文献报道的一致;构建了CaMv35S启动子驱动hCu,Zn—SOD基因的植物表达载体PBICuSOD;用三亲交配法将重组质粒PBICuSOD转化到农杆菌LBA4404,转化马铃薯得到转基因植株。PCR和Southern—blot分析证明,hCu,Zn—SOD基因巳整合到马铃薯基因组中。Western blot分析表明,hCu,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1066.6U/g(湿重,下同),块茎的总酶活为10llU/g。

       

      Abstract: 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).

       

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