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Characterization and molecular cloning of the PCF8775 CS5 antigen from an enterotoxigenic Escherichia coli 0115:H40 isolated in Central Australia
Author(s) -
Heuzenroeder M. W.,
Neal B. L.,
Thomas C. J.,
Halter R.,
Manning P. A.
Publication year - 1989
Publication title -
molecular microbiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.857
H-Index - 247
eISSN - 1365-2958
pISSN - 0950-382X
DOI - 10.1111/j.1365-2958.1989.tb00175.x
Subject(s) - biology , protein subunit , pilin , escherichia coli , gene , cloning (programming) , molecular cloning , enterotoxigenic escherichia coli , coding region , peptide sequence , oligonucleotide , microbiology and biotechnology , biochemistry , pilus , enterotoxin , computer science , programming language
Summary The genes determining the biosynthesis of the colonization factor CS5 have been cloned from Escherichia coli 0115:H40:PCF8775 isolated during an outbreak of diarrhoea among aboriginal children in Central Australia. Electron microscopy has shown purified CS5 to be of semi‐rigid fimbrial type. NH 2 ‐terminal analysis has shown the CS5 determinant to be distinct from other fimbriae, although there is some conservation of certain residues. Expression in minicells of the cloned fimbrial genes encoded on pPM1312 has shown that proteins of 70 and 46.5 kD which co‐purify with the 23kD major fimbrial subunit protein are also co‐expressed along with proteins of 45, 31, 17 and 14kD. The major CS5 subunit is synthesized in precursor form (approximately 26 kD). A synthetic oligonucleotide to the NH2‐terminal amino acid coding sequence of the purified protein has been used in Southern hybridization analyses to define the region on pPM1312 encoding the structural gene for the major pilin subunit