粪肠球菌介导的日本血吸虫重组疫苗的构建、鉴定及表达  

Construction,identification and expression of recombinant Schistosoma japonicum vaccine mediated by Enterococcus faecalis

作　　者：李文桂[1] 欧兴坤 何爱琳 Li Wengui;Ou Xingkun;He Ailin(Institute of Infections and Parasitic Diseases,the First Afiliated Hospital,Chonging Medical University,Chongging 400016,China)

机构地区：[1]重庆医科大学附属第一医院传染病寄生虫病研究所,重庆400016

出　　处：《中华地方病学杂志》2023年第3期173-177,共5页Chinese Journal of Endemiology

基　　金：国家自然科学基金(30801052、30671835、30500423、30200239)。

摘　　要：目的:构建粪肠球菌(Efs)介导的日本血吸虫(Sj)重组疫苗(rEfs-Sj26GST疫苗),并研究Sj26GST-GST融合蛋白在重组疫苗中的表达情况。方法:将pGEX-Sj26GST重组质粒电穿孔转化Efs ATCC47077株感受态,构建rEfs-Sj26GST疫苗,提取质粒进行PCR鉴定;经异丙基-β-D-硫代半乳糖苷(IPTG)诱导表达后,用十二烷基硫酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和蛋白质印迹法(Western blot)对表达产物进行分析和鉴定。结果:经PCR鉴定,扩增出大小约676 bp的片段,与Sj26GST扩增片段长度相符。经SDS-PAGE分析,得到相对分子质量约52 × 10 3的条带,与Sj26GST-GST融合蛋白条带相符。Western blot结果显示,rEfs-Sj26GST疫苗表达的Sj26GST-GST融合蛋白可被Sj感染者血清特异性识别。 结论:成功构建了rEfs-Sj26GST疫苗,Sj感染者血清可特异性识别重组疫苗表达的Sj26GST-GST融合蛋白。Objective To construct a recombinant vaccine of Schistosoma japonicum(Sj)mediated by Enterococcus faecalis(Efs,rEfs-Sj26GST vaccine),and to study the expression of Sj26GST-GST fusion protein in the recombinant vaccine.Methods The recombinant plasmid pGEX-Sj26GST was transformed into the susceptible strain Efs ATCC47077 by electroporation to construct rEfs-Sj26GST vaccine,and the plasmid was extracted for PCR identification.After induction of expression with isopropyl-beta-D-thiogalactopyranoside(IPTG),the products were analyzed and identified by sodium dodecyl sulfate-polyacrylamide gel electrophoresis(SDS-PAGE)and Western blot.Results After PCR identification,a 676 bp fragment was amplified,which was consistent with the length of Sj26GST amplification fragment.SDS-PAGE analysis showed that the relative molecular mass was 52×10',which was consistent with the band of Sj26GST-GST fusion protein.Western blot results showed that the Sj26GST-GST fusion protein expressed by rEfs-Sj26GST vaccine could be specifically recognized by the serum of Sj infected patients.Conclusion The rEfs-Sj26GST vaccine is successfully constructed,and the Sj26GST-CST fusion protein expressed by recombinant vaccine can be specifically recognized by the serum of Sj infected patients.

关 键 词：粪肠球菌 日本血吸虫 重组疫苗 Sj26GST-GST融合蛋白 

分 类 号：R392-33[医药卫生—免疫学]