ESCHERICHIA-COLI CS31A FIMBRIAE - MOLECULAR-CLONING, EXPRESSION AND HOMOLOGY WITH THE K88-DETERMINANT

被引:24
作者
MARTIN, C
BOEUF, C
BOUSQUET, F
机构
[1] Laboratoire de Microbiologie, Institut National de la Recherche Agronomique, Centre de Clermont-Ferrand-Theix
关键词
BACTERIAL FIMBRIAE; ESCHERICHIA-COLI; MOLECULAR HYBRIDIZATION; ADHESIN;
D O I
10.1016/0882-4010(91)90108-M
中图分类号
R392 [医学免疫学]; Q939.91 [免疫学];
学科分类号
100102 ;
摘要
CS31A is a plasmid-encoded K88-related fimbrial antigen. A Sau3AI library was constructed from p31A, a 180 kb CS31A encoding plasmid, in the pSUP202 vector. Bacterial recombinant clones expressing CS31A were isolated. A 8.5 kb EcoRI-HindIII DNA fragment from one of them was subcloned in pBR322 and pHSG575 vectors, leading to pAG315 and pEH524 recombinant plasmids respectively. Escherichia coli harboring pAG315 or pEH524 expressed CS31A fimbrial antigens on their cell surface. Analysis of these plasmids in minicells showed that at least seven mature polypeptides were encoded by the EcoRI-HindIII DNA fragment, with apparent molecular masses of 76000, 54000, 30000, 29000, 28000, 15500 and 13500 daltons respectively. The genetic organization of the CS31A gene cluster was determined and showed to be similar to that of the K88 operon. The nucleotide sequence homology between CS31A and K88 determinants was investigated by Southern blot hybridization at high stringency. This indicated that extensive nucleotide sequence homology exists throughout both gene clusters except for the subunit structural genes. © 1991.
引用
收藏
页码:429 / 442
页数:14
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