Degenerate PCR primers for the amplification of fragments from genes encoding response regulators from a range of pathogenic bacteria.
Many bacterial responses to environmental stimuli are mediated by response regulators which coordinately regulate genes involved in particular adaptive responses. Degenerate oligonucleotide primers were used to amplify by the polymerase chain reaction (PCR), fragments from genes encoding eleven novel response regulators. Sequence and phylogenetic analysis revealed that phoB, phoP and creB gene fragments had been amplified from Yersinia enterocolitica and Yersinia pseudotuberculosis, and that a creB sequence had been amplified from Campylobacter jejuni. Four amplified fragments from C. jejuni, Listeria monocytogenes, Mycobacterium tuberculosis and Escherichia coli clearly came from response regulator genes, but were not closely related to any of the known genes. Mutagenesis of the newly identified genes should allow us to determine their function and the genes under their control.
Wren
B W
BW
Department of Medical Microbiology, St. Bartholomew's Hospital Medical College, West Smithfield, London, UK.
Colby
S M
SM
Cubberley
R R
RR
Pallen
M J
MJ
D000595:Amino Acid Sequence; D001419:Bacteria / Q000235:genetics / Q000472:pathogenicity; D001425:Bacterial Outer Membrane Proteins / Q000235:genetics; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000235:genetics*; D005798:Genes, Bacterial; D005809:Genes, Regulator; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes; D010802:Phylogeny; D016133:Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid; D013045:Species Specificity
D001425:Bacterial Outer Membrane Proteins; D004269:DNA, Bacterial; D015345:Oligonucleotide Probes