豆豉纤溶酶基因的克隆及其在大肠杆菌中的表达  

Cloning of the Douchi Fibrinolytic Enzyme Gene and its Expression in Escherichi coli

作　　者：陆云华[1] 张新[1] 

机构地区：[1]宜春学院化学与生物工程学院,江西宜春336000

出　　处：《安徽农业科学》2007年第23期7104-7104,7107,共2页Journal of Anhui Agricultural Sciences

基　　金：江西省教育厅科研项目(编号:2007-314)

摘　　要：根据已发表的豆豉纤溶酶基因DNA序列(GeneBank AY720895.2),设计并合成了一对引物,应用PCR技术以筛选的来自豆豉的产纤溶酶的芽孢杆菌的总DNA为模板,扩增出了豆豉纤溶酶基因,并将其克隆到pMD18-T载体上,进行了序列测定,测序结果经Genetool序列分析表明该基因长1 089 bp,编码363个氨基酸,与已发表Brevibacillus laterosporus豆豉纤溶酶基因序列在核苷酸水平上与所发表的序列有98%的同源性;在氨基酸水平上与已发表的芽孢杆菌豆豉纤溶酶同源性为100%。并将该基因克隆到大肠杆菌表达载体pET22b上,在大肠杆菌中实现了高效表达。According to the published gene sequence of douchi fibrinolytic enzyme （DFE） from GeneBank AY720895.2, primers were designed to amplify DFE gene by PCR. The PCR product was cloned to pMD18-T and sequenced. The cloned DFE gene comprised 1089 nucleotides and encoded a polypeptide of 363 amino acids. Compared with the published gene sequence in GeneBank AY720895.2, the DFE gene showed 98 % identity on nucleotides level and 100 % identity on amino acids level. DFE gene was also cloned to the expression vector pET22b and overexpressed in E.coli.

关 键 词：豆豉纤溶酶 基因克隆 高效表达 

分 类 号：Q78[生物学—分子生物学]