鼠防御素基因Crp4在大肠杆菌中的表达及活性检测  

Prokaryotic Expression of Mouse Defensin Crp4 in Escherichia coli and Its Activity Detection

作　　者：陈宝玉[1] 宋德光[1] 宋斯伟[1] 查云峰 邓旭明[1] 曾凡勤[1] 

机构地区：[1]吉林大学畜牧兽医学院,长春130062 [2]广东省动物防疫监督总所,广州510230

出　　处：《吉林农业大学学报》2006年第5期577-580,590,共5页Journal of Jilin Agricultural University

基　　金：国家"924"项目(2004AA002060)

摘　　要：通过基因工程技术,以鼠防御素基因Crp4为研究对象,从鼠回肠末端克隆了鼠防御素Crp4基因的成熟编码序列(约为102 bp),将其与另外2个鼠防御素基因的氨基酸序列进行比对,发现同源性分别高达97.8%和97.1%。将该序列克隆到原核表达载体pET-28a中,构建了重组质粒pET-Crp4,转化DE3感受态并用IPTG进行诱导表达。SDS-PAGE和Wersiernblot结果表明,该基因在pET-28a中成功表达。体外抑菌试验证实,重组Crp4蛋白对金黄色葡萄球菌、李斯特菌、大肠杆菌等具有一定的抑菌活性。Using genetic engineering technique, mouse defeusin Crp4 with its length 102 bp is cloned from mouse ileum. Through sequence analysis of Crp4 with other two mouse defeusius reported, we found their homogeneity is up to 97.8 % and 97.1% . Then the target DNA fragment was cloned into pET-28a vector, and the recombinant vectors were transformed into E. coli DE3, so the expression was induced based on the optimal values of the IPTG, concentration incubation temperature and induction time determined in the previous section. The expressed proteius were analyzed by result of SDS-PAGE and Wertem blot. The recombinant Crp4 fusion proteins possess the antimicrobial activity to staphylococcus aureus, E. coli in the assay of drug susceptibility by the method of agar diffusion. So we can further study biological characteristics of defeusin and its medical value.

关 键 词：防御素 Crp4 大肠杆菌 原核表达 生物学活性 

分 类 号：Q786[生物学—分子生物学] S852.61[农业科学—基础兽医学]