Cloning and organization of seven arginine biosynthesis genes from Neisseria gonorrhoeae

A genomic library for Neisseria gonorrhoeae, constructed in the A cloning vector EMBL4. was screened for clones carrying arginine biosynthesis genes by complementation of Escherichia coli mutants. Clones complementing defects in argA, argB, argE, argG, argIF, carA and carB were isolated. An E. coli...

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Bibliographic Details
Published in:Journal of Bacteriology 1989-03, Vol.171 (3), p.1644-1651
Main Authors: Picard, F.J, Dillon, J.R
Format: Article
Language:English
Subjects:
acetylglutamate synthetase
acetylornithine deacetylase
acyltransferases
amide hydrolases
amino acid metabolism
arga gene
argb gene
arge gene
argif gene
arginine
Arginine - biosynthesis
Bacteriology
Biological and medical sciences
cara gene
carb gene
carbamoyl-phosphate synthase (glutamine-hydrolysing)
cloning
Cloning, Molecular
Escherichia coli - genetics
Fundamental and applied biological sciences. Psychology
Genes
Genes, Bacterial
genetic complementation
Genetic Complementation Test
Metabolism. Enzymes
Microbiology
mutants
Neisseria
Neisseria gonorrhoeae
Neisseria gonorrhoeae - enzymology
Neisseria gonorrhoeae - genetics
Nucleic Acid Hybridization
ornithine acetyltransferase
Plasmids
structural genes
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Summary:A genomic library for Neisseria gonorrhoeae, constructed in the A cloning vector EMBL4. was screened for clones carrying arginine biosynthesis genes by complementation of Escherichia coli mutants. Clones complementing defects in argA, argB, argE, argG, argIF, carA and carB were isolated. An E. coli defective in the acetylornithine deacetylase gene (argE) was complemented by the ornithine acetyltransferase gene (argJ) from N. gonorrhoeae. This heterologous complementation is reported for the first time. The carAB operon from E. coli hybridized with the gonococcal clones that carried carA or carB genes under conditions of high stringency, detecting 80% or greater similarity and showing that the nucleotide sequence of the carbamoylphosphate synthetase genes is very similar in these two organisms. Under these conditions for hybridization, the gonococcal clones carrying argB or argF genes did not hybridize with plasmids containing the corresponding E. coli gene. Cocomplementation experiments established gene linkage between carA and carB. Clones complementing a gene defect in argE were also able to complement an argA mutation. This suggests that the enzyme ornithine acetyltransferase from N. gonorrhoeae (encoded by argJ) may be able to complement both argA and argE mutations in E. coli. The arginine biosynthesis genes in N. gonorrhoeae appear to be scattered as in members of the family Pseudomonadaceae.
ISSN:0021-9193
1098-5530
1067-8832
DOI:10.1128/JB.171.3.1644-1651.1989