大肠杆菌铜锌超氧化物歧化酶基因的克隆与序列测定

›› 2010, Vol. 37 ›› Issue (5): 75-78.

大肠杆菌铜锌超氧化物歧化酶基因的克隆与序列测定

袁伟，唐善虎，章轶锋，陈诺，胡慧玲

（西南民族大学生命科学与技术学院，成都 610041）

收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-05-20 发布日期:2010-05-20
通讯作者: 唐善虎

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

YUAN Wei，TANG Shan-hu，ZHANG Yi-feng,CHEN Nuo,HU Hui-ling

(College of Life Science & Technology, Southwestern University for Nationalities, Chengdu 610041， China)

Received:1900-01-01 Revised:1900-01-01 Online:2010-05-20 Published:2010-05-20
Contact: TANG Shan-hu

摘要/Abstract

摘要： 试验以E.coli MG1655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.coli DH5α感受态细胞，筛选阳性克隆，提取质粒进行Sac I和Kpn I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。

关键词: 铜锌超氧化物歧化酶; 分子克隆; 序列测定

Abstract: 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.

Key words: superoxide dismutase; cloning; sequence analysis

中图分类号:

袁伟;唐善虎;章轶锋;陈诺;胡慧玲. 大肠杆菌铜锌超氧化物歧化酶基因的克隆与序列测定[J]. , 2010, 37(5): 75-78.

YUAN Wei;TANG Shan-hu;ZHANG Yi-feng;CHEN Nuo;HU Hui-ling. Cloning and Sequencing of the Escherichia coli Copper-Zinc Superoxide Dismutase[J]. , 2010, 37(5): 75-78.

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