Summary
In vitro recombination techniques were used to clone the E. coli metJBLF gene cluster in a plasmid vector. Several chimeric plasmids were obtained, analyzed by restriction mapping and characterized genetically. The combined results establish that the met gene cluster is contained on an approximately 5.6 kilobase segment of bacterial DNA with metL between metB and metF. The origin of metL was localized precisely by its DNA sequence and its transcription direction was established.
Similar content being viewed by others
References
Bachmann BJ, Low KB (1980) Linkage map of Escherichia coli K12, ed 6. Bacteriol Rev 44:1–56
Birnboim HC, Doly J (1979) A rapid alkaline extraction procedure for screening recombinant plasmid DNA. Nucl Acids Res 7:1513–1523
Bolivar F, Rodriguez RL, Greene PJ, Betlach ML, Heyneker HL, Boyer HW (1977) Construction and characterization of new cloning vehicles; II. A multipurpose cloning systeme. Gene 2:95–113
Clewell DB (1972) Nature of ColE1 plasmid replication in Escherichia coli in the presence of chloramphenicol. J Bacteriol 110:667–676
Cohn M, Cohen GN, Monod J (1953) L'effet inhibiteur spécifique de la méthionine dans la formation de la méthionine-synthase chez Escherichia coli. CR Acad Sci Paris 236:746–748
Cossart P, Katinka M, Yaniv M, Saint-Girons I, Cohen GN (1979) Construction and expression of a hybrid plasmid containing the Escherichia coli thrA and thrB genes. Mol Gen Genet 175:39–44
Dautry-Varsat A, Sibilli-Weill L, Cohen GN (1977) Subunit structure of the methionine repressible aspartokinase II-homoserine dehydrogenase II from Escherichia coli K12. Eur J Biochem 76:1–6
Greene RC, Krueger JH (1982) Localization of the metF gene in Escherichia coli: Construction and properties of a recombinant λmet transducing bacteriophage. Bacteriol J (submitted)
Haseltine B, Müller-Hill B (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor NY pp 319–325
Johnson JR, Greene RC, Krueger JH (1977) Isolation and characterization of specialized lambda transducing bacteriophage carrying the metBJF methionine gene cluster. J Bacteriol 131:795–800
Kahn M, Kolter R, Thomas C, Figurski D, Meyer R, Remaut E, Helinski DR (1979) Plasmid cloning vehicles derived from plasmids ColE1, F, R6K and RK2. Method Enzymol 68:268–280
Katinka M, Cossart P, Sibilli L, Saint-Girons I, Chalvignac MA, Le Bras G, Cohen GN, Yaniv M (1980) Nucleotide sequence of the thrA gene of Escherichia coli. Proc Natl Acad Sci USA 77:5730–5733
Katz L, Kingsbury DT, Helinski DR (1973) Stimulation by cyclic adenosine monophosphate of plasmid deoxyribonucleic acid replication and catabolite repression of the plasmid deoxyribonucleic acid protein complex. J Bacteriol 114:577–591
Kingsbury DT, Helinski DR (1973) Temperature-sensitive mutants for the replication of plasmids in Escherichia coli: Requirement for deoxyribonucleic acid polymerase I in the replication of the plasmid ColE1. J Bacteriol 114:1116–1124
Konrad EB (1969) Ph D Thesis, Harvard University, Cambridge
Krueger JH, Johnson JR, Greene RC, Dresser M (1981) Structural studies of lambda transducing bacteriophage carrying bacterial deoxyribonucleic acid from the metBJLF region of the Escherichia coli chromosome. J Bacteriol 147:612–621
Maniatis T, Jeffery A, Sande H van de (1975) Chain length determination of small double and single stranded DNA molecules by polyacrylamide gel electrophoresis. Biochemistry 14:3787–3794
Maxam AM, Gilbert W (1977) A new method for sequencing DNA. Proc Natl Acad Sci USA 74:560–564
McDonell 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
Miller JH (1972) Experiments in molecular genetics. Cold Spring, Cold Spring Harbor Laboratory, Harbor NY, p 433
Norgard MV, Emigholz K, Monahan JJ (1979) Increased amplification of pBR322 plasmid deoxyribonucleic acid in Escherichia coli K12 strains RR1 and χ 1776 grown in the presence of high concentrations of nucleoside. J Bacteriol 138:270–272
Ohtsubo E, Lee HJ, Deonier RC, Davidson N Electron microscope heteroduplex studied of sequence relations among the plasmids of Escherichia coli. VI. Mapping of F14 sequences homologous to ϕ80dmetBJF and ϕ80dargECBH bacteriophages. J Mol Biol 89:599–618
Patte JC, Le Bras G, Cohen GN (1967) Regulation by methionine of the synthesis of a third aspartokinase and of a second homoserine dehydrogenase in Escherichia coli K12. Biochim Biophys Acta 136:245–257
Peacock A, Dingman CW (1967) Resolution of multiple ribonucleic acid species by polyacrylamide gel electrophoresis. Biochemistry 6:1818–1827
Press R, Glansdorff N, Miner P, de Vries J, Kadner R, Maas WK (1971) Isolation of transducing particles of ϕ80 bacteriophage that carry different regions of the Escherichia coli genome. Proc Natl Acad Sci USA 68:795–798
Radloff R, Bauer W, Vinograd J (1967) A dyebuoyant density method for the detection and isolation of closed circular duplex DNA: the closed circular DNA in HELA cells. Proc Natl Acad Sci USA 57:1514–1521
Rowbury RJ (1965) Resistance to norleucine and control of methionine synthesis in Escherichia coli. Nature 206:962–963
So M, Gill R, Falkow S (1976) The generation of ColE1-Apr cloning vehicle which allows detection of inserted DNA. Mol Gen Genet 142:239–249
Su CH, Greene RC (1971) Regulation of methionine biosynthesis in Escherichia coli: Mapping of the metJ locus and properties of a metJ +/metJ− diploid. Proc Natl Acad Sci USA 68:367–371
Tanaka T, Weisblum B (1975) Construction of a colicin E1-R factor composite plasmid in vitro: Means for amplification of deoxyribonucleic acid. J Bacteriol 121:345–362
Thèze J, Margarita D, Cohen GN, Borne F, Patte JC (1974) Mapping of the structural genes of the three aspartokinases and the two homoserine dehydrogenases of Escherichia coli K12. J Bacteriol 117:133–143
Tiollais P, Rambach A, Buc H (1974) Large scale purification of a bacterial gene. FEBS Lett, 48:96–100
Wijesundera S, Woods DD (1953) The effect of growth on a medium containing methionine on the synthesis of this amino acid by Bacterium coli. Biochem J 55:viii
Zakin MM, Garel JR, Dautry-Varsat, Cohen GN, Boulot G (1978) Detection of the homology among proteins by immunochemical cross-reactivity between denatured antigens. Application to the threonine and methionine regulated aspartokinases-homoserine dehydrogenases from E. coli K12. Biochemistry 17:4318–4323
Author information
Authors and Affiliations
Additional information
Communicated by G.O.'Donovan
Rights and permissions
About this article
Cite this article
Zakin, M.M., Greene, R.C., Dautry-Varsat, A. et al. Construction and physical mapping of plasmids containing the metJBLF gene cluster of E. coli K12. Molec. Gen. Genet. 187, 101–106 (1982). https://doi.org/10.1007/BF00384390
Received:
Issue Date:
DOI: https://doi.org/10.1007/BF00384390