大肠杆菌铜锌超氧化物歧化酶基因的克隆与序列测定  

Cloning and Sequencing of the Escherichia coli Copper-Zinc Superoxide Dismutase

作　　者：袁伟[1] 唐善虎[1] 章轶锋[1] 陈诺[1] 胡慧玲[1] 

机构地区：[1]西南民族大学生命科学与技术学院,成都610041

出　　处：《中国畜牧兽医》2010年第5期75-78,共4页China Animal Husbandry & Veterinary Medicine

基　　金：西南民族大学人才引进项目(234610)

摘　　要：试验以E.coliMG1655的基因组DNA为模板,通过PCR技术扩增sodC基因,PCR产物经纯化回收后克隆至pMD18-T载体中,转化E.coliDH5α感受态细胞,筛选阳性克隆,提取质粒进行SacI和KpnI酶切及PCR扩增鉴定,并对sodC基因片段进行序列测定。试验成功克隆了sodC基因,获得的基因与报道的sodC基因序列同源性达到99.6%,为进一步构建重组表达载体奠定了基础。Using chromosome DNA of Escherichia coli MG1655 as template,sodC gene was amplified by PCR and purified by purification kit in this study. The PCR product of sodC gene was cloned into pMD18-T vector and transformed into DH5α competent cells. The recombinant plasmid containing sodC gene was selected and the inserted sodC gene was identified by cleaved with restriction endonuclease Sac I and Kpn I,and following by PCR amplification and sequencing. The results showed that the whole sodC gene was cloned successfully. Sequence analysis indicated that sodC gene displayed 99.6% nucleotide identities with the published sequences by GenBank. This research proved a foundation for the construction of recombinant expression vector of sodC gene.

关 键 词：铜锌超氧化物歧化酶 分子克隆 序列测定 

分 类 号：Q78[生物学—分子生物学]