河流弧菌B7-1羧肽酶基因的克隆与功能鉴定  

Cloning and functional identification of Vibrio fluvialis B7-1 carboxylpeptidase gene

作　　者：韦松松 谢晓东 陈帅君 黄海东 杨红澎 吴疆 Wei Songsong;Xie Xiaodong;Chen Shuaijun;Huang Haidong;Yang Hongpeng;Wu Jiang(College of Agronomy and Resource Environment,Tianjin Agricultural University,Tianjin 300392,China)

机构地区：[1]天津农学院农学与资源环境学院,天津300392

出　　处：《天津农学院学报》2022年第2期18-21,26,共5页Journal of Tianjin Agricultural University

基　　金：国家级大学生创新创业训练计划项目(202010061011);天津市科技计划项目(20ZYCGSN00390)。

摘　　要：羧肽酶是一类外切酶,它能专一性地从肽链C末端逐一切下单个氨基酸,在生物体中多以酶原的形式存在。近年来无论是在生物界还是在医学行业,羧肽酶越来越引起人们的关注。本研究从细菌Vibrio fluvialis中提取全基因组,设计上下游引物进行编码羧肽酶基因的克隆,将扩增的羧肽酶基因转入到TOP10大肠杆菌中进行质粒扩增,然后在BL-21大肠杆菌中进行目的蛋白的表达,通过对其表达的羧肽酶进行定性定量分析,可以确定重组的包含Vibrio fluvialis的羧肽酶基因在大肠杆菌中成功表达。Carboxypeptidase is a type of exonuclease that can specifically cleave individual amino acids one by one from the c-terminus of a peptide chain. In recent years, carboxypeptidase has attracted more and more attention in both biological and medical industry. In this study, the whole genome was obtained from bacterial Vibrio fluvialis, and an upstream primer and a downstream primer were designed to amplify the target gene encoding carboxypeptidase. The amplified target gene was transferred into TOP10 E.coli for recombinant amplification, and then the target protein was expressed in E.coli BL-21.Qualitative and quantitative analysis of the carboxypeptidase expression revealed that the recombinant strain containing the carboxypeptidase gene of V. fluvialis had strong capability of expressing carboxypeptidase and strong expressed enzyme activity.

关 键 词：基因 羧肽酶 酶活 Folin-酚法 

分 类 号：Q786[生物学—分子生物学]