保加利亚乳杆菌N-乙酰胞壁质酶的原核表达和鉴定  

Prokaryotic Expression and Identification of Lactobacillus bulgaricus N-acetylmuramidase

作　　者：张玉玉[1] 苏文政[1] 马建军[1] 黄保华[1] 蒋慧 吴家强[1] 王可[1] Zhang Yuyu;Su Wenzheng;Ma Jianjun;Huang Baohua;Jiang Hui;Wu Jiaqiang;Wang Ke(Institute of Animal Husbandry and Veterinary Medicine, Shandong Academy ofAgricultural Sciences/Shandong Key Laboratory of Animal Disease Control and Breeding, Jinan 250100, China;Shandong Institute for Food and Drug Control, Jinan 250101, China)

机构地区：[1]山东省农业科学院畜牧兽医研究所/山东省畜禽疫病防治与繁育重点实验室,山东济南250100 [2]山东省食品药品检验研究院,山东济南250101

出　　处：《山东农业科学》2018年第5期14-17,共4页Shandong Agricultural Sciences

基　　金：山东省农业科学院青年科研基金项目(2015YQN52);山东省农业重大应用技术创新项目;济南市金种子企业关键产品提升计划项目(201502069)

摘　　要：为研究保加利亚乳杆菌(LB)N-乙酰胞壁质酶(Mur)的分子生物学特性,根据已发表的保加利亚乳杆菌核苷酸序列,设计并合成一对特异性引物,PCR扩增LB MN株不含跨膜区域序列的mur基因,片段回收后克隆至pMD19-T载体并进行鉴定,再将其亚克隆于原核表达载体pET-30a(+)中构建重组表达质粒pET-30a(+)-mur,转化大肠杆菌BL21(DE3),IPTG诱导重组蛋白表达。SDS-PAGE和Western blot分析显示重组Mur蛋白分子量约为21 kD,能在大肠杆菌中高效表达。In order to study the molecular properties of the N-acetylmuramidase fromLactobacillus bulgaricus,a pair of special primers were designed and synthesized according to the nucleotide sequence of LB mur gene in Gen Bank. The mur gene without transmembrane sequence was amplified from the genome DNA of LB MN strain and cloned into pMD19-T vector. The gene was sequenced and cloned into the prokaryotic expression vector pET-30 a（＋）. The recombinant plasmid pET-30 a（＋）-mur was transformed into competent BL21（DE3） and the expression of the recombinant protein was induced with IPTG. The SDS-PAGE and Western blot showed the molecular weight of the recombinant protein mur was approximately 21 kD and its expression was efficient in Escherichia coli.

关 键 词：保加利亚乳杆菌 N-乙酰胞壁质酶 原核表达 鉴定 

分 类 号：S879.1[农业科学—畜牧兽医] TS252.1[轻工技术与工程—农产品加工及贮藏工程]