Bacillus sp．nov SK006中纤维蛋白溶解酶的克隆及其在大肠杆菌中的表达  

Cloning of Fibrinolytic Enzyme from Bacillus sp.nov SK006 and Its Expression in Escherichia coli

作　　者：秦芸桦[1,2] 沐万孟[1] 江波[1] 

机构地区：[1]江南大学食品科学与技术国家重点实验室,江苏无锡214122 [2]a江南大学食品学院,江苏无锡214122

出　　处：《食品与发酵工业》2011年第9期22-24,共3页Food and Fermentation Industries

基　　金：国家自然科学基金(20976073);江南大学内自主科研项目(JUSRP21107)

摘　　要：利用PCR技术从Bacillus sp.nov SK006中扩增得到纤维蛋白溶解酶(Fibrinolitic enzymes,FE)基因的DNA片段,将其连接到pMD-T载体上,经测序后克隆至载体PET-22b(+),转化至E.coli BL21(DE3)中,得到的重组菌能高效表达FE。诱导表达后,SDS-PAGE显示,重组FE分子量约为28 ku。A DNA encoding fibrinolytic enzyme（FE） was amplified from Bacillus sp. nov SK006 using PCR. Sequencing showed open reading frame is composed of 828 bp encoding a putative protein of 276 amino acid residues. The cDNA was cloned into an expression vector pET- 22b（ ＋ ） and expressed in Escherichia coli BL21 （DE3）. The recombinant protein was purified to electrophoretical homogeneity by affinity chromatography using Chelating Sepharose Fast Flow resin. The purified recombinant FE had an approximate molecular weight of 28 ku by SDS-PAGE analvsis.

关 键 词：虾酱 纤维蛋白溶解酶 克隆 表达 

分 类 号：Q78[生物学—分子生物学]