人铜锌超氧化物歧化酶(hCu,Zn-SOD)基因在马铃薯中的表达

Expression of Human Copper,Zinc-superoxide Dismutase(hCu,Zn-SOD)Gene in Potato

目的 构建转基因马铃薯植株 ,以便大规模生产人铜锌超氧化物歧化酶 (hCu ,Zn SOD)。方法 采用RT PCR技术从人肝总RNA中分离扩增了hCu Zn SOD基因 ,并将其重组到植物表达载体pPSCuSOD中 ,用三亲交配法将重组质粒pPSCuSOD转化到农杆菌LBA44 0 4,再用农杆菌介导转化马铃薯。结果 经PCR和PCR Southern分析证明hCu Zn SOD基因已整合到马铃薯基因组中。Westernblot分析证明 ,hCu Zn SOD基因已在马铃薯植株中得到表达。NBT光还原法检测转基因马铃薯块茎中的SOD总酶活性为 192 0U g .FW。

Objective To construct a recombinant strain of potato for the large scale production of human copper,zinc superoxide dismutase(hCu,Zn SOD).Methods Isolate and amplify hCu,Zn SOD gene from the total human liver RNA,and clone into a plant expression vector pPSCuSOD.Introduce the recombinant plasmid pPSCuSOD into Agrobacterium tumefaciens LBA4404 and transform to potato.Results PCR and PCR Southern analysis proved that hCu,Zn SOD gene was integrated into potato genome.Western blot showed that hCu,Zn SOD gene was highly expressed in patato.Conclusion The SOD activity of transgenic potato,detected by NBT photoreduction,was 1920U/g.FW.