Summary
Genes have been cloned from Salmonella typhimurium which when present on the multicopy plasmid pBR322 in the E. coli strain NT31 confer a Gua+ phenotype on this strain. NT31 is a purE gpt double mutant and it was expected that a Gua+ phenotype could be conferred on it by the cloning of either gpt or purE. It was, however found that in addition to these two loci the molecular cloning of another gene, which has been identified as hpt, in pBR322 confers a Gua+ phenotype on NT31. This result is explained by the overproduction of the hpt gene product, hypoxanthine phosphoribosyl transferase, which compensates for the lack of the gpt product guanine-xanthine phosphoribosyl transferase. Restriction analysis of the three loci, gpt, hpt and purE is also presented.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- Kb:
-
kilobase pairs
- Tc:
-
tetracyline
- m.o.i:
-
multiplicity of infection
- 8AG:
-
8-azaguanine
References
Birnboim HC, Doly J (1979) A rapid extraction procedure for screening recombinant plasmid DNA. Nucl Acids Res 7:1513–1523
Bolivar F, Rodriguez RL, Greene PJ, Betlach MC, Heyneker HL, Boyer HW (1977) Construction and characterization of new cloning vehicles II: A multipurpose cloning system. Gene 2:95–113
Cantwell BA, McConnell DJ (1983) Molecular cloning and expression of a Bacillus subtilis β-glucanase in Escherichia coli. Gene 23:211–219
Chou JY, Martin RG (1972) Purine phosphoribosyl-transferases of Salmonella typhimurium. J Bacteriol 112:910–916
Cohen SN, Chang ACY, Hsu L (1972) Nonchromosomal antibiotic resistance in bacteria: genetic transformation of Escherichia coli by R-factor DNA. Proc Natl Acad Sci USA 69:2110–2114
Davis RW, Botstein D, Roth JR (1980) Advanced bacterial genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Gots JS (1971) Regulation of purine and pyrimidine metabolism. In: Vogel HJ (ed) Metabolic regulation, vol V. Academic Press Inc, New York, p 225–255
Gots JS, Benson CE, Shumas SR (1972) Genetic separation of hypoxanthine and guanine-xanthine phosphoribosyltransferase activities by deletion mutations in Salmonella typhimurium. J Bacteriol 112:910–916
Gots JS, Benson CE, Jochimsen B, Koduri KR (1977) Microbial models and regulatory elements in the control of purine metabolism. Ciba Foundation Symp 48:23–41
Hammer-Jespersen K, Munch-Petersen A, Nygaard P, Schwartz M (1971) Induction of enzymes involved in the catabolism of deoxyribonucleosides and ribonucleosides in Escherichia coli. Eur J Biochem 19:533–538
Hochstadt-Ozer J (1971) Regulation of purine utilization in bacteria 1. Purification of adenine phosphoribosyltransferase from Escherichia coli K12 and control of activity of nucleotides. J Biol Chem 246:5294–5303
Hoppe I, Roth JR (1974) Specialized transducing phages derived from Salmonella phage P22. Genetics 76:633–654
Itikawa H, Demerec M (1968) Salmonella typhimurium proline mutants. J Bacteriol 95:1189–1190
Jessop AP (1972) A specialised transducing phage of P22 for which the ability to form plaques is associated with transduction of the proAB region. Mol Gen Genet 114:214–222
Jessop AP (1976) Specialized transducing phages derived from phage P22 that carry the proAb region of the host, Salmonella typhimurium: Genetic evidence for their structure and mode of transduction. Genetics 83:459–475
Jochimsen B, Nygaard P, Vestergaard T (1975) Location on the chromosome of Escherichia coli of genes governing purine metabolism. Mol Gen Genet 143:85–91
Krenitsky TA, Neil SM, Miller RL (1970) Guanine and xanthine phosphoribosyl-transfer activities of Lactobacillus casei and Escherichia coli. J Biol Chem 245:2605–2611
Lowry OH, Rosebrough NJ, Farr AL, Randall RJ (1951) Protein measurement with the Folin phenol reagent. J Biol Chem 193:265–275
Maniatis T, Fritsch EF, Sambrook J (1982) Molecular cloning — a Laboratory Manual. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Martin WR, Yang RR (1972) Inosine and guanosine phosphoribo-syltransferase in Escherichia coli. Biochem Biophys Res Commun 48:1641–1648
McConnell DJ, Bonner J (1972) Preparation of highly purified ribonucleic acid polymerase; separation from polynucleotide phosphorylase and polyphosphate kinase. Biochemistry 11:4329–4336
McDonnell MW, Simon MN, Studier FW (1977) Analysis of restriction fragments of T7 DNA and determination of molecular weights by electrophoresis in neutral and alkaline gels. J Mol Biol 110:119–146
Miyake T, Demerec M (1960) Proline mutants of Salmonella typhimurium. Genetics 45:755–762
Sanderson KE, Roth JR (1983) Linkage map of Salmonella typhimurium. Edition VI Microbiol Rev 47:410–553
Sanger F, Coulson AR, Hong GH, Hill DF, Petersen GB (1982) Nucleotide sequence of bacteriophage λ DNA. J Mol Biol 162:729–773
Smith HO, Levine M (1965) Gene order in prophage P22. Virology 27:229–231
Smith HO, Levine M (1967) A phage P22 gene controlling integration of prophage. Virology 31:207–216
Smith-Keary PF (1960) A suppressor of leucineless in Salmonella typhimurium. Heredity 14:61–71
Smith-Keary PF (1966) Restricted transduction of prophage P22 in Salmonella typhimurium. Genet Res 8:73–82
Smith SM, Stocker BAD (1966) Mapping of prophage P22 in Salmonella typhimurium. Virology 28:413–419
Vogel HJ, Bonner DM (1956) Acetylornithase of Escherichia coli: partial purification and some properties. J Biol Chem 218:97–106
Author information
Authors and Affiliations
Additional information
Communicated by A. Böck
Rights and permissions
About this article
Cite this article
O'Reilly, C., Turner, P.D., Smith-Keary, P.F. et al. Molecular cloning of genes involved in purine biosynthetic and salvage pathways of Salmonella typhimurium . Mol Gen Genet 196, 152–157 (1984). https://doi.org/10.1007/BF00334108
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00334108