铜绿假单胞菌弹性蛋白酶基因的扩增与克隆

Amplification and cloning of elastase coding sequence from pseudomonas aeruginosa

目的PCR扩增及克隆铜绿假单胞菌的弹性蛋白酶基因。方法从铜绿假单胞菌培养物中提取染色体DNA,PCR扩增弹性蛋白酶基因成熟蛋白编码区,AT克隆于pMD18T载体中,对重组质粒进行酶切与测序鉴定。结果从高GC含量的铜绿假单胞菌染色体DNA中扩增到弹性蛋白酶基因并进行了克隆与鉴定。结论本研究成功克隆到铜绿假单胞菌弹性蛋白酶基因,为实现活性弹性蛋白酶的高效表达奠定了基础。

Objective To amplify and clone the elastase coding sequence from pseudomonas aeruginosa.Methods Genomic DNA was extracted from pseudomonas aeruginosa culture, and the elastase coding sequence was amplified by using PCR technique, then linked to pMD 18-T vector and transformed to E. coli JM109. The recombinant plamid was identified by Bam HI and Kpn I digestion and sequencing.Results The elastase coding sequence of pseudomonas aeruginosa was amplified successfully and been cloned and identified.Conclusion The cloning of the elastase coding sequence of pseudomonas aeruginosa provides the basis for the high level expression of its coding protein.