Open AccessMolecular cloning of the S‐layer protein gene of Campylobacter rectus ATCC 33238
Author(s) -
Miyamoto M.,
Kobayashi Y.,
Kokeguchi S.,
Ohta H.,
Kurihara H.,
Fukui K.,
Murayama Y.
Publication year - 1994
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.1574-6968.1994.tb06668.x
Subject(s) - microbiology and biotechnology , s layer , biology , gene , hspa2 , plasmid , molecular cloning , genetics , gene expression , peptide sequence
A genomic DNA library of Campylobacter rectus (Wolinella recta) ATCC 33238 was constructed using bacteriophage lambda EMBL3 as a vector. One clone expressing the S‐layer protein was identified immunologically with the antiserum to the S‐layer protein of C. rectus ATCC 33238. Western immunoblotting using monoclonal antibodies directed against the S‐layer protein showed a single blot of recombinant protein at 150 kDa, suggesting that this clone contained the entire coding region of the S‐layer protein gene. Additionally, the S‐layer protein gene was subcloned into plasmid vector pUC18. Southern hybridization revealed that the S‐layer protein gene was present on the chromosome of C. rectus as a single‐copy gene, and that there were minor heterogeneities among the S‐layer protein genes of clinical isolates. Moreover, a spontaneous stable mutant strain, which has no S‐layer expression, also conserved the full‐length gene of the S‐layer protein.