假单胞菌ZL13邻苯二酚2,3-双加氧酶基因的克隆表达

Cloning of a new catechol 2,3-dioxygenase gene from Pseudomonas sp.ZL13 and its expression in the E.coli BL21

摘要:

寻找新型高效石油降解菌,并研究其相关基因一直是石油降解领域的热点.本文以细菌Pseudomonas sp.ZL13的降解性质粒pZL-1的DNA为模板,通过PCR扩增的方法进行基因克隆,得到邻苯二酚2,3-双加氧酶基因.序列分析结果表明,该基因大小为924bp,编码307个氨基酸;序列同源性比较结果显示,该基因与荧光假单胞菌(Pseudomonas fluorescens)和恶臭假单胞菌(Pseudomonas putida)的基因序列的相似性较高,处于同一分支.将目的基因连接到pGEX-4T-2表达载体上,在E.coli BL21中成功表达,转化子具有原油降解能力.

Abstract:

Rearches about screening oil-degrading bacteria and their genes involed in the degrading procedure are always important.In this paper,rocatechol-2,3 pyrocatechol gene was cloned from plasmid pZL-1 by PCR method.This gene is 924 bp long and encoding apeptide of 307 amino acids.After multiple sequence alignment,it is found that the c23O gene from P.putida.ZL13 is highly similar with the c23O gene from P.fluorescens and P.putida.The pyrocatechol-2,3 pyrocatechol gene was ligated to pGEX-4T-2 and transformed in E.coli BL21.The results show that this gene can be successfully expressed.