微小隐孢子虫LCCL结构域基因的克隆及原核表达  

Cloning and Prokaryotic Expression of LCCL Domain Gene of Cryptosporidium parvum

作　　者：宁良辰[1] 张颖[1] 赵权[1] 

机构地区：[1]吉林农业大学动物科技学院,吉林长春130118

出　　处：《动物医学进展》2013年第3期68-71,共4页Progress In Veterinary Medicine

摘　　要：为探讨微小隐孢子虫的入侵机制,克隆、表达了微小隐孢子虫LCCL结构域基因。从已构建的微小隐孢子虫cDNA文库中,采用PCR随机扩增LCCL结构域基因。与Pmd-18-T载体连接后,挑取阳性重组子测序分析。用基因重组技术将LCCL结构域基因克隆入原核表达载体pGEX-4T-1中,构建LCCL结构域基因的重组原核表达载体pGEX-4T-1-LCCL。重组质粒经酶切、测序鉴定后转入大肠埃希菌BL21中,IPTG诱导表达目的蛋白,并用SDS-PAGE和Western blot进行鉴定。最终得到了在大肠埃希菌中高效表达的重组蛋白,分子质量大小约为37ku,并且具有反应原性,为进一步研究微小隐孢子虫的入侵机制奠定了基础。To explore the mechanism of Cryptosporidium parvurn invasion, the LCCL-domain gene of Cryptosporidium parvum was cloned and expressed. Total DNA was extracted from cDNA library of Cryptosporidium parvurn and the DNA fragment encoding mature peptide of LCCL-d0main gene was am- plified by PCR. Coding sequence identified by DNA sequencing was cloned into the expression vector pGEX-4T-1 to construct prokaryotic expression vector. The E. coli BL21 containing recombinant plasmid was induced by IPTG to express fusion protein. SDS-PAGE and Western blot were performed to identify the recombinant protein. The result of Western blot demonstrated that the expressed protein had reaetogenieity. The results laid the foundation for the further study of Cryptosporidium parvum invasion mechanism.

关 键 词：微小隐孢子虫 LCCL结构域基因 克隆 原核表达 

分 类 号：S852.723[农业科学—基础兽医学]