人铜锌超氧化物歧化酶的基因工程研究——Ⅰ.人铜锌超氧化物歧化酶基因的合成与克隆

Chemical Synthesis and Cloning of a Gene for Human Cu/Zn Superoxide Dismutase

本文报道人铜锌超氧化物歧化酶(Cu/Zn superoxide Dismutase, SOD)基因的合成与克隆。采用大肠杆菌偏爱密码子,按入Cu/Zu SOD一级结构顺序编码。基因全长495个碱基对,合成时,分成26个基因片段分别合成。采用分级退火的方法,在T4 DNA连接酶作用下,将基因片段组装成完整的基因,经序列分析证明,合成的基因顺序与设计一致。

The SOD gene, which encodes human Cu/Zn superoxide dismutase, has been chemically synthesised and cloned. The synthetic gene contains the structure genei a BamH Ⅰ site and start signal at 5' end; a Hind Ⅲ site and stop signal at 3 ' end. Computer programming was applied to the design of optimized segments for the synthesis of SOD gene, 26 segments were synthesized by the solid phase phosphoramidite method, with lengths ranging from 26 to 44 nucleotides. The whole gene was constructed by enzymatic ligation of all the oligonucleotides. The sequence of the synthetic SOD gene was proved correct by DNA sequencing analysis.