猕猴溶组织内阿米巴凝集素hgl基因LC3段的克隆及原核表达  

Cloning and prokaryotic expression of Gal/GalNAc lectin hgl LC3 gene of Entamoeba histolytica isolated from Macaca mulatta

作　　者：余星明[1] 姜波[2] 杨光友[2] 王强[1] 余建秋[1] 牛李丽[1] 古小彬[2] 邓家波[1] 程安春[2] 毕凤均[3] 陈维刚[1] 

机构地区：[1]成都动物园,四川成都610081 [2]四川农业大学动物医学院,四川雅安625014 [3]四川农业大学实验动物中心国家实验猕猴种源基地,四川雅安625014

出　　处：《中国兽医科学》2010年第2期139-143,共5页Chinese Veterinary Science

基　　金：成都大熊猫繁育研究基金项目(06-02);教育部"长江学者和创新团队发展计划"创新团队项目(IRT0848);国家科技基础条件平台项目(2004DKA30660)

摘　　要：为了研究动物溶组织内阿米巴基因工程疫苗,以川西猕猴溶组织内阿米巴分离株中提取的总基因组DNA为模板,扩增了hgl基因抗原区LC3段,将其重组于原核表达载体pET-32b(+)中,构建了pET-32b(+)-hglLC3表达载体,转化至大肠杆菌BL21中,经IPTG诱导表达融合蛋白,获得了1200bp的hgl基因LC3编码区片段;SDS-PAGE分析结果显示,重组表达质粒产生了大小为66ku的目的条带,与预期的结果一致;Western-blot分析结果显示,重组蛋白与兔超免疫血清呈阳性反应,表明,该抗原可作为动物溶组织内阿米巴疫苗的候选抗原。To develop genetic engineering vaccine against Entamoeba histolytica infection in animals,a LC3 segment of hgl gene of E.histolytica was amplified from total genomic DNA of E.histolytica from Macaca mulatta by RT-PCR,and then subcloned into the pET-32b（＋） vector.The recombinant vector pET-32b（＋）-hglLC3 was transformed into Escherichia coli BL21 for expression with IPTG induction.The LC3 segment of hgl gene of E.histolytica was 1 200 bp.SDS-PAGE analysis indicated that the expressed fusion protein in recombinant E.coli was approximate 66 ku in molecular weight.Western-blot analysis showed that the fusion protein could be recognized by the hyperimmune sera from the rabbits inoculated with E.histolytica proteins,indicating that the recombinant protein expressed from the LC3 segment of hgl gene could be used as a candidate antigen for development of effective vaccines against E.histolytica infection in animals.

关 键 词：川西猕猴 溶组织内阿米巴 凝集素hgl基因 LC3段 原核表达 

分 类 号：S852.721[农业科学—基础兽医学]