摘要
用基因重组技术获得的高效表达肠毒素大肠杆菌定居日子抗原Ⅰ（ＣＦＡ／Ⅰ）的重组克隆，通过匀浆破碎、硫酸铵分级盐析部分纯化，再经超速离心及ＤＥＡＥ－ＳｅｐｈａｄｅｘＡ５０柱层析，得到了ＣＰＡ／Ⅰ的纯化样品，得率约为０．９ｍｇ／ｇ湿菌。聚丙烯酰胺凝胶电泳（ＳＤＳ－ＰＡＧＥ）显示一条蛋白质带，其分子量为１５ｋＤａ，与天然ＣＦＡ／Ⅰ相同。免疫扩散和蛋白质印迹试验证明，纯化的重组克隆ＣＦＡ／Ⅰ与天然ＣＦＡ／Ⅰ具有相同的抗原性及免疫原性。并在电镜下观察了ＣＦＡ／Ⅰ的菌毛形态。
The recombinant clone E. coli C 600(pZLH88)can efficiently
express the colonization factor antigen 1(CFA/Ⅰ)of enterotoxigenic Escherichia coli.The fimbrial
CFA/Ⅰ was purified and charerized.The initial puri-fication step was the release of these
fimbriae from the bacterial cells by homogenization. The essential purificationsteps were
ammonium suifate fractionation,ultracentrifugation and DEAE-Sephadex A 50 column
chromatography. The final yield of CFA/Ⅰprotein was 0.9mg of antigen per g(wet weight)of
bacteria. On SDS-PAGE, this prepa-ration produced a single polypeptide subunit band of 15 kDa
which was the same with the native CFA/Ⅰ protein.Purified CFA/Ⅰ is a fimbrial molecule 7.0 nm
in diameter.Analysis of purified CFA/Ⅰby immunodiffusion andWestern immunoblot showed
that this specific antiserum against purified CFA/Ⅰ antigen did react with the nativeCFA/Ⅰ
subunits.
出处
《军事医学科学院院刊》
CSCD
北大核心
1994年第2期127-132,共6页
Bulletin of the Academy of Military Medical Sciences
关键词
大肠杆菌
定居因子抗原
纯化
ETEC
enterotoxin E. coli
colonization factor antigcn Ⅰ
recombinant clone
purification 1 Institute of Mcdical Sciences,Xinjiang Military Region,PLA2 Department of
Pathology,Agriculture and Livestock Husbandry University, PLA
