霍乱弧菌EL－Tor35A3菌株外膜蛋白主区带的纯化  

PURIFICATION OF MAJOR BAND OUTER MEMBRANE PROTEIN OF VIBRIO CHOLERAE EL-TOR

作　　者：林厚怡[1] 陈慎宝[1] 宋兰珍 丁如宁[1] 沈凤新 

机构地区：[1]南京医科大学生化教研室,南京医科大学微生物教研室

出　　处：《微生物学通报》1995年第6期358-361,共4页Microbiology China

基　　金：国家自然科学基金

摘　　要：选用霍乱弧菌ＥＬ－Ｔｏｒ生物型３５Ａ３菌株（血清稻叶型）以ＥＤＴＡ－溶菌酶法与超速离心法提取其外膜蛋白，并将聚丙烯酰胺凝胶电泳后一条蛋白主区带分离。ＳＤＳ－聚丙烯酰胺凝胶电泳测得该蛋白主区带的分子量为２５ｋＤ，免疫电泳显示一条线，免疫双扩试验表明，不同的霍乱弧菌菌株均有该蛋白抗原存在。Vibrio cholerae,like other Gram-negative bacteria,has an outer membranecontaining lipopolysaccharide(LPS)and outer membrane protein(OMP).Both OMP andLPS are essential in the making of cholera vaccine. This Investigation is aimed to analyzethe OMP of the vibrio cholerae helping to get an in-depth knowledge the chemical compo-sition of the bacterral cell envelope in the making of cholera vaccine.Vibrio cholerae EL-Tor 35A3,BH43,WJ2 and Classical 569B were used in this study.lsolation of OMP was accomplished by treating the cell envelope with 1% EDTA and 0.5%lysozyme for shaking at 37℃ 2h followed by centrifugation at 105000g for 1h.The majorband of OMP was isolated with polylarcylamide electrophoresis and analyzed by electropho-resis which contained SDS-PAGE and immuno-electrophoresis.The results suggest that allthese strains contained a major protein antigen band of molecular weight 25kD and have across-reacting the protein antigen located in the outer membrane.This finding,the authorssupposed,may be a light in design of a cholera vaccine,but this study is primary.It isnecessary that the further study is making to do.

关 键 词：霍乱弧菌 外膜蛋白 主区带 纯化 

分 类 号：R378.3[医药卫生—病原生物学]