多房棘球绦虫Em14-3-3抗原编码基因在BCG中的表达  

Expression of Echinococcus multilocularis 14-3-3 antigen encoding gene in BCG

作　　者：王鸿[1] 李文桂[1] 

机构地区：[1]重庆医科大学传染病寄生虫病研究所,重庆400016

出　　处：《重庆医科大学学报》2006年第3期299-301,共3页Journal of Chongqing Medical University

基　　金：教育部重点项目(205131);重庆市教委科研基金(KJ050313)

摘　　要：目的:研究多房棘球绦虫Em14-3-3抗原编码基因在BCG中的表达。方法:将重组质粒pBCG-Em14-3-3用电穿孔法导入BCG构建rBCG,将含有重组子的细菌培养至对数生长期,在收菌前3天每天45℃热诱导30min,然后对表达产物作SDS-PAGE及免疫印迹分析。结果:多房棘球绦虫pBCG-Em14-3-3在BCG中成功表达了Em14-3-3重组蛋白,在相对分子量为27KDa处可见明显的表达蛋白带,其表达量占菌体总蛋白量的11%。结论:多房棘球绦虫pBCG-Em14-3-3能在BCG中高效表达,且能与鼠血清抗体发生特异结合,提示rBCG-Em14-3-3疫苗表达的Em14-3-3重组蛋白具有特异的抗原性。Objective： To study the expression of Echinococcus multilocularis 14-3-3 antigen encoding gene in Bacillus Calmette-Guerin （BCG）. Methods： The recombinant plasmid pBCG-Em 14-3-3 was introduced into BCG by electroporation and the bacteriawith pBCG-Em14-3-3 were cultured to logarithm growth period, then induced daily by heating at 45℃ for 30min in the last three days.The expression products were analyzed by SDS-PAGE and Western blot. Results： The Em14-3-3 recombinant protein was successfullyexpressed by pBCG-Em14-3-3 in BCG as an obvious band of 27kDa by SDS-PAGE, which accounted for 11% of total bacterial proteinin BCG. Conclusion： The pBCG- Em14-3-3 could be expressed efficiently in BCG and be combined with specific antibody in mice sera, which suggested that the Em 14-3-3 recombinant protein expressed by rBCG- Em 14-3-3 vaccine was of specific antigenicity.

关 键 词：多房棘球绦虫 重组Em14-3-3蛋白 BCG 基因表达 

分 类 号：R383.33[医药卫生—医学寄生虫学]