Summary
The technique developed by Rosenfeld and Brenchley [J Bacteriol 144, 848–851 (1980)] has been used to introduce Mu d1 (Apr lac) into Salmonella typhimurium for purposes of constructing pyr::lac fusion strains. A stable pyrB::lac fusion mutant was subsequently derived and used for the genetic characterization of the pyrB gene. The direction of transcription of pyrB was determined to be counterclockwise on the S. typhimurium linkage map and argI was shown to be located clockwise of pyrB. Mutants altered in the regulation of expression of pyrB were isolated and two of the isolates chosen for further study were tentatively categorized as promoter or operator mutants.
Similar content being viewed by others
References
Abdelal ATH, Griego E, Ingraham JL (1976) Arginine sensitive phenotype of mutations in pyrA of Salmonella typhimurium: role of ornithine carbamyltransferase in the assembly of mutant carbamylphosphate synthetase. J Bacteriol 128:105–113
Bachmann BJ, Low KB (1980) Linkage map of Escherichia coli K-12, edition 6. Microbiol Rev 44:1–56
Back KJC, Woods DD (1953) Studies with a strain of Bacterium coli requiring citrulline and a pyrimidine for growth. Biochem J 55:xii-xiii
Beckwith JR (1970) Lac: the genetic system. In: Beckwith JR, Zipser D (eds) The lactose operon. Cold Spring Harbor Laboratory, New York, pp 5–26
Bukhari AI (1975) Reversal of mutator phage Mu integration. J Mol Biol 96:87–99
Casadaban MJ, Cohen SN (1979) Lactose genes fused to exogenous promoters in one step using a Mu-lac bacteriophage: in vivo probe for transcriptional control sequences. Proc Natl Acad Sci USA 76:4530–4533
Chumley FG, Menzel R, Roth JR (1978) Hfr formation directed by Tn10. Genetics 91:639–655
Csonka LN, Howe MM, Ingraham JL, Pierson III LS, Turnbough CL (1981) Infection of Salmonella typhimurium with coliphage Mu d1 (Apr lac): construction of pyr::lac gene fusions. J Bacteriol 145:299–305
Faelen M, Mergeay M, Gerits J, Toussaint A, Lefèbvre N (1981) Genetic mapping of a mutation conferring sensitivity to bacteriophage Mu in Salmonella typhimurium LT2. J Bacteriol 146:914–919
Ferro-Luzzi Ames G, Biek DP, Spudich EN (1978) Duplications of histidine transport genes in Salmonella typhimurium and their use for the selection of deletion mutants. J Bacteriol 136:1094–1108
Glansdorff N, Dambly C, Palchaudhuri S, Crabeel M, Pierard A, Halleux P (1976) Isolation and heterduplex mapping of a lambda transducing bacteriophage carrying the structural genes for carbamoylphosphate synthase: regulation of enzyme synthesis in Escherichia coli K-12 lysogens. J Bacteriol 127:302–308
Ingraham JL, Neuhard J (1972) Cold-sensitive mutants of Salmonella typhimurium defective in uridine monophosphate kinase (pyrH). J Biol Chem 247:6259–6265
Jensen KF (1979) Apparent involvement of purines in the control of expression of Salmonella typhimurium pyr genes: analysis of a leaky guaB mutant resistant to pyrimidine analogs. J Bacteriol 138:731–738
Justesen J, Neuhard J (1975) pyrR identical to pyrH in Salmonella typhimurium: control of expression of the pyr genes. J Bacteriol 123:851–854
Kelln RA, Kinahan JJ, Foltermann KF, O'Donovan GA (1975) Pyrimidine biosynthetic enzymes of Salmonella typhimurium, repressed specifically by growth in the presence of cytidine. J Bacteriol 124:764–744
Kleckner N, Chan RK, Tye B-K, Botstein D (1975) Mutagenesis by insertion of a drug-resistance element carrying an inverted repetition. J Mol Biol 97:561–575
Kleckner N, Roth J, Botstein D (1977) Genetic engineering in vivo using translocatable drug-resistance elements. New methods in bacterial genetics. J Mol Biol 116:125–159
Lee, JH, Heffernan L, Wilcox G (1980) Isolation of ara-lac gene fusions in Salmonella typhimurium LT2 by using transducing bacteriophage Mu d (Apr lac). J Bacteriol 143:1325–1331
Lee J-H, Al-Zarban S, Wilcox G (1981) Genetic characterization of the araE gene in Salmonella typhimurium LT2. J Bacteriol 146:298–304
Levine RA, Taylor MW (1982) Regulation of purE transcription in a purE::lac fusion strain of Escherichia coli. J Bacteriol 149:1041–1049
MacPhee DG, Beazer MR (1973) Mutants of Salmonella typhimurium deficient in DNA polymerase I: detection by their failure to produce colicin E1. Mol Gen Genet 127:229–240
Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, New York
Miller JR, Kline BC (1979) Biochemical characterization of nonintegrated plasmid-folded chromosome complexes: sex factor F and the Escherichia coli nucleoid. J Bacteriol 137:885–890
Mitchell A, Finch LR (1979) Enzymes of pyrimidine metabolism in Mycoplasma mycoides subsp. mycoides. J Bacteriol 137:1073–1080
O'Donovan GA, Gerhart JC (1972) Isolation and partial characterization of regulatory mutants of the pyrimidine pathway in Salmonella typhimurium. J Bacteriol 109:1085–1096
Piérard A, Glansdorff N, Yashphe J (1976) Mutations affecting uridine monophosphate pyrophysphorylase or the argR gene in Escherichia coli. Effects on carbamoyl phosphate and pyrimidine biosynthesis and on uracil uptake. Mol Gen Genet 118:235–245
Rosenfeld SA, Brenchley JE (1980) Bacteriophage P1 as a vehicle for Mu mutagenesis of Salmonella typhimurium. J Bacteriol 144:848–851
Roth JR (1970) Genetic techniques in studies of bacterial metabolism. Meth Enzymol 27A:3–35
Sanderson KE, Hartman PE (1978) Linkage map of Salmonella typhimurium, edition V. Microbiol Rev 42:471–519
Schwartz M, Neuhard J (1975) Control of expression of the pyr genes in Salmonella typhimurium: effects of variations in uridine and cytidine nucleotide pools. J Bacteriol 121:814–822
Susskind M, Botstein D (1978) Molecular genetics of bacteriophage P22. Microbiol Rev 42:385–413
Syvanen JM, Roth JR (1972) Structural genes for ornithine transcarbamylase in Salmonella typhimurium and Escherichia coli K-12. J Bacteriol 110:66–70
Syvanen JM, Roth JR (1973) Structural genes of catalytic and regulatory subunits of aspartate transcarbamylase. J Mol Biol 76:363–378
Williams JC, Lee CE, Wild JR (1980) Genetic and biochemical characterization of distinct transport systems for uracil, uridine and cytidine in Salmonella typhimurium. Mol Gen Genet 178:121–130
Williams JC, O'Donovan GA (1973) Repression of enzyme synthesis of the pyrimidine pathway in Salmonella typhimurium. J Bacteriol 115:1071–1076
Zak VL, Kelln RA (1978) 5-Fluoroorotate-resistant mutants of Salmonella typhimurium Can J Microbiol 24:1339–1345
Zak VL, Kelln RA (1981) A Salmonella typhimurium mutant dependent upon carbamyl aspartate for resistance to 5-fluorouracil is specifically affected in ubiquinone biosynthesis. J Bacteriol 145:1095–1098
Author information
Authors and Affiliations
Additional information
Communicated by G.A. O'Donovan
Rights and permissions
About this article
Cite this article
Michaels, G., Kelln, R.A. Construction and use of pyr::lac fusion strains to study regulation of pyrimidine biosynthesis in Salmonella typhimurium . Molec. Gen. Genet. 189, 463–470 (1983). https://doi.org/10.1007/BF00325910
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00325910