【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。更多还原

【Abstract】 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.更多还原