Cloning and characterization of the catalase gene of Neisseria gonorrhoeae: use of the gonococcus as a host organism for recombinant DNA.
The structural gene for the catalase of Neisseria gonorrhoeae was cloned into a Kat- strain of that organism by using a recombinant vector derived from one of the beta-lactamase-specifying plasmids found in that organism. The kat gene was then successfully subcloned into both pUC8 and pGB2, transformed into Escherichia coli, and shown to complement the E. coli katE mutants UM2 and UMRl. The gene was subsequently mutagenized and returned to the gonococcus to generate a Kat- strain that was phenotypically identical to the strain originally used to clone the gene. The sequence of the gene and the derived amino acid sequence showed that the gonococcal kat gene closely resembles the hktE gene of Haemophilus influenzae. The sequence of the promoter region of the gonococcal kat gene is unusual and may explain the extremely high, loosely regulated expression of the gene.
Johnson
S R
SR
Division of AIDS, STD, and TB Laboratory Research, National Center for Infectious Diseases, Centers for Disease Control and Prevention, Atlanta, Georgia 30333, USA.
Steiner
B M
BM
Perkins
G H
GH
D000595:Amino Acid Sequence; D001483:Base Sequence; D002374:Catalase / Q000235:genetics*; D003001:Cloning, Molecular; D004274:DNA, Recombinant / Q000235:genetics*; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005798:Genes, Bacterial; D005816:Genetic Complementation Test; D006193:Haemophilus influenzae / Q000201:enzymology / Q000235:genetics; D008969:Molecular Sequence Data; D009154:Mutation; D009344:Neisseria gonorrhoeae / Q000201:enzymology* / Q000235:genetics*; D010641:Phenotype; D010957:Plasmids / Q000235:genetics; D015183:Restriction Mapping
D004274:DNA, Recombinant; D002374:Catalase