Rapid Cloning and Expression of Glutaryl-7-Aminocephalosporanic Acid Acylase Genes from Soil Samples

Rapid Cloning and Expression of Glutaryl-7-Aminocephalosporanic Acid Acylase Genes from

Soil Samples

LUO Hui;YU Huimin;LI Qiang;SHEN Zhongyao

【期刊名称】《清华大学学报（英文版）》 【年(卷),期】2005(010)005

【摘要】A polymerase chain reaction (PCR)-based strategy was developed to rapidly obtain the gene encoding for an industrially important enzyme, glutaryl-7-aminocephalosporanic acid (GL-7-ACA) acylase. Different soil samples were cultured with a Pseudomonas selective medium to enrich specific microorganisms, and then the genomic DNA was extracted to serve as PCR templates. PCR primers for GL-7-ACA acylase gene amplification were designed on the basis of bioinformatics searches and analyses. The method was used to successfully amplify three GL-7-ACA acylase genes from different soil samples. The GL-7-ACA acylase genes were then cloned and overexpressed in Escherichia coli with a relatively high level of 266 unit·L-1.

【总页数】6页(529-534)

【关键词】glutaryl-7-aminocephalosporanic acid acylase;polymerase chain reaction;soil;Pseudomonas

【作者】LUO Hui;YU Huimin;LI Qiang;SHEN Zhongyao