Open AccessIdentification and molecular cloning of a 70 kDa species‐specific antigen common to Clostridium difficile
Author(s) -
Wren Brendan W.,
Mullany Peter P.,
Clayton Christopher L.,
Tabaqchali Soad
Publication year - 1988
Publication title -
fems microbiology letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.899
H-Index - 151
eISSN - 1574-6968
pISSN - 0378-1097
DOI - 10.1111/j.1574-6968.1988.tb02370.x
Subject(s) - clostridium difficile , identification (biology) , cloning (programming) , biology , antigen , microbiology and biotechnology , molecular cloning , computational biology , genetics , gene , peptide sequence , antibiotics , computer science , programming language , botany
Three common antigens (CB 1, 2 and 3), characteristic of Clostridium difficile species were identified by immunoblot analysis using homologous and heterologous rabbit antisera, raised against whole cells from 9 distinct strains of C. difficile . A gene library of C. difficile genomic DNA was constructed in Escherichia coli by cloning in Sau 3A‐cleaved clostridial DNA fragments into the bacteriophage vector λEMBL3. Out of 3000 plaques screened using the whole cell antisera, 27 clones were positively identified. One of these clones, designated λCd21, expressed high levels of an antigen which could be immunologically identified using whole cell antisera against the 9 C. difficile strains. Antiserum raised against the clone λCd21 identified a 70 kDa antigen (previously named CB1) as demonstrated by immunoblot analysis. Monospecific antiserum against λCd21 recognises the 70 kDa antigen in all 97 strains of C. difficile derived from worldwide sources and does not cross‐react with 17 strains from 13 other clostridial species.