日本血吸虫Mf1和胞质氨基肽酶基因的亚克隆表达与免疫保护效果测定  

Subcloning,expression and vaccination studies with Schistosoma japonicum Mf1 and cytosol aminopeptidase genes

作　　者：王庆林[1] 易新元[1] 曾宪芳[1] 罗新松 何永康 彭兴华[3] Larry Me Reynolds 

机构地区：[1]中南大学湘雅医学院寄生虫学研究室 [2]湖南省寄生虫病研究所 [3]中南大学湘雅医学院分子生物学中心 [4]New England Biolabs

出　　处：《中国人兽共患病杂志》2003年第2期38-41,共4页Chinese Journal of Zoonoses

基　　金：家九五科技攻关项目 (编号 :96 -A2 3- 0 6 -k0 4 )

摘　　要：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 。Aim To subclone and express of Schistosoma japonicum(Sj) Mf1 gene and cytosol aminopeptidase(CA) gene in E coli and to evaluate the immunoprotective effect of these recombinant molecules Methods The cDNA of Sj Mf1 gene and SjCA gene were then used to transform E coli strain ER2688 Expressionwas induced by IPTG The expressed proteins were vaccinated to mice to evaluate the immunoprotective effect The mice in vaccination groups were immunized subcutaneously with 50 μg recombinant protein emulsified with Freund's complete adjuvant(FCA),and those in control groups were injected FCA or PBS at week 0,2,6 The mice were challenged by Sj cercariae two weeks after the final injection,and were sacrificed 30 days after challenge,worm burden were counted Results By IPTG inducing,fusion proteins SjGST-Mf1 and SjGST-CA were high expressed in E Coli in insoluble form,the inclusion bodies The reduction in worm burden in vaccination group of SjGST-Mf1 and of SjGST-CA was 39 15% and 26 57%,respectively Conclusion SjMf1 gene and SjCA gene could be high expressed in E coli after subcloned into pGEX-5X-3 Fusion proteins Sj GST-Mf1 and Sj GST-CA can significantly induce anti-infection immunity

关 键 词：日本血吸虫 Mf1 胞质氨基肽酶基因 亚克隆表达 免疫保护 测定 

分 类 号：R383.24[医药卫生—医学寄生虫学]