摘要
目的:构建xylE基因高表达菌株,为快速检测芳香类化合物污染提供可能。方法和结果:通过PCR扩增,将位于质粒pTG402上的xylE基因进行扩增,克隆于pUC118N和pUC119N上,经双向测序,证明PCR过程中并未发生突变,然后进一步克隆于高表达载体pJLA503,转化E.coliTG1,经42℃诱导,获得了高表达的基因产物,表达量占全菌总蛋白的34.2%。结论:xylE基因在大肠杆菌中获得了高表达。
Objective:High level expression of xylE gene in E.coli is available to detect aromatic compound environment pollution.Methods and Results:The xylE gene which codes for the catechol 2,3 dioxygenase(CatO2ase) was amplified from the recombinant plasmid pTG402 by using PCR technique and was subcloned onto pUC118N and pUC119N.The single stranded recombinant phage DNA from the transformed E.coli MV1184 cells was used for sequencing.The sequence of xylE gene was proved to be the same as reported.The gene was then subcloned onto the high expression plasmid pJLA503,and its expression amount was about 34.2% in total bacterial proteins.Conclusion:xylE gene is highly expressed in host E.coli TG1.
出处
《第二军医大学学报》
CSCD
北大核心
1996年第4期306-312,共7页
Academic Journal of Second Military Medical University
基金
国家自然科学基金