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目的 在大肠杆菌中高效表达日本血吸虫 (Schistosomajaponicum ,Sj)Mf1蛋白和胞质氨基肽酶 (cytosolaminopeptidase ,CA) ,并对表达产物进行免疫保护效果测定。 方法 将SjMf1基因和SjCA基因亚克隆至 pGEX - 5X - 3载体 ,转化入感受态大肠杆菌ER2 688,在IPTG诱导下进行表达 ,表达产物免疫小鼠 ,免疫用抗原剂量为 50 μg/次 /鼠 ,免疫 3次后进行攻击感染 ,观察诱导产生的减虫效果。结果 在IPTG诱导下 ,表达载体中的SjGST基因与重组的SjMf1基因和SjCA基因在大肠杆菌中获得了高效表达 ,形成SjGST -Mf1和SjGST -CA融虫蛋白 ,用这两种融合蛋白免疫小鼠 ,分别诱导产生了 39 1 5 %和 2 6 57%的减虫率。结论 SjMf1和SjCA基因亚克隆至pGEX - 5X - 3载体后可在大肠杆菌中高效表达 ,表达产物能诱导小鼠产生一定程度的抗Sj保护性免疫力
Objective To express Mf1 protein and cytosolaminopeptidase (CA) of Schistosoma japonicum (Sj) efficiently in Escherichia coli (E.coli), and to determine the immunoprotective effect of the expressed product. Methods SjMf1 gene and SjCA gene were subcloned into pGEX - 5X - 3 vector and transformed into competent E.coli ER2688 for expression under the induction of IPTG. The expressed product was used to immunize mice with the antigen at a dose of 50 μg / , Immunized 3 times after the attack infection, observed the effect of induced de-worming. Results Under the induction of IPTG, the SjGST gene, the recombinant SjMf1 gene and the SjCA gene in the expression vector were highly expressed in Escherichia coli to form the SjGST-Mf1 and SjGST-CA melt proteins. The two fusion proteins were used to immunize mice , Respectively, induced 39 1 5% and 2 6 57% reduction of worm rates. Conclusion The SjMf1 and SjCA genes were subcloned into pGEX - 5X - 3 vector and expressed efficiently in E. coli. The expressed product of SjMf1 and SjCA gene induced some degree of anti - Sj protective immunity in mice