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Molecular cloning and expression of Clostridium difficile toxin A in Escherichia coli K 12
Author(s) -
Wren Brendan W.,
Clayton Christopher L.,
Mullany Peter P.,
Tabaqchali Soad
Publication year - 1987
Publication title -
febs letters
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.593
H-Index - 257
eISSN - 1873-3468
pISSN - 0014-5793
DOI - 10.1016/0014-5793(87)81135-3
Subject(s) - escherichia coli , microbiology and biotechnology , toxin , cloning (programming) , clostridium difficile , clostridium , molecular cloning , clostridium difficile toxin b , microbial toxins , clostridium difficile toxin a , chemistry , enterotoxin , bacteria , biology , gene , gene expression , genetics , biochemistry , antibiotics , computer science , programming language
Clostridium difficile toxin A was purified to homogeneity and was used to raise monospecific antiserum in rabbits. A gene bank of C . difficile DNA in Escherichia coli was constructed by cloning Sau 3A‐cleaved clostridial DNA fragments into the bacteriophage vector λEMBL3. Out of 4500 plaques screened with antitoxin A, 9 clones were positively identified. One of these clones λtA5 expressed a 235 kDa protein which exhibited a cytotonic effect on Chinese hamster ovary cells, and had the ability to haemagglutinate rabbit erythrocytes, both properties characteristic of toxin A. The size of the λtA5 insert DNA was 14.3 kb.