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Journal of Bacteriology logoLink to Journal of Bacteriology
. 1987 Apr;169(4):1454–1459. doi: 10.1128/jb.169.4.1454-1459.1987

Molecular cloning, characterization, and chromosomal localization of dapF, the Escherichia coli gene for diaminopimelate epimerase.

C Richaud, W Higgins, D Mengin-Lecreulx, P Stragier
PMCID: PMC211967  PMID: 3031013

Abstract

The Escherichia coli dapF gene was isolated from a cosmid library as a result of screening for clones overproducing diaminopimelate epimerase. Insertional mutagenesis was performed on the cloned dapF gene with a mini-Mu transposon, leading to chloramphenicol resistance. One of these insertions was transferred onto the chromosome by a double-recombination event, allowing us to obtain a dapF mutant. This mutant accumulated large amounts of LL-diaminopimelate, confirming the blockage in the step catalyzed by the dapF product, but did not require meso-diaminopimelate for growth. The dapF gene was localized in the 85-min region of the E. coli chromosome between cya and uvrD.

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Selected References

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  1. ANTIA M., HOARE D. S., WORK E. The stereoisomers of alpha epsilon-diaminopimelic acid. III. Properties and distribution of diaminopimelic acid racemase, an enzyme causing interconversion of the LL and meso isomers. Biochem J. 1957 Mar;65(3):448–459. doi: 10.1042/bj0650448. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Aiba H., Mori K., Tanaka M., Ooi T., Roy A., Danchin A. The complete nucleotide sequence of the adenylate cyclase gene of Escherichia coli. Nucleic Acids Res. 1984 Dec 21;12(24):9427–9440. doi: 10.1093/nar/12.24.9427. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bachmann B. J. Pedigrees of some mutant strains of Escherichia coli K-12. Bacteriol Rev. 1972 Dec;36(4):525–557. doi: 10.1128/br.36.4.525-557.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Boy E., Patte J. C. Multivalent repression of aspartic semialdehyde dehydrogenase in Escherichia coli K-12. J Bacteriol. 1972 Oct;112(1):84–92. doi: 10.1128/jb.112.1.84-92.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Bradford M. M. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem. 1976 May 7;72:248–254. doi: 10.1016/0003-2697(76)90527-3. [DOI] [PubMed] [Google Scholar]
  6. Bukhari A. I., Taylor A. L. Genetic analysis of diaminopimelic acid- and lysine-requiring mutants of Escherichia coli. J Bacteriol. 1971 Mar;105(3):844–854. doi: 10.1128/jb.105.3.844-854.1971. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Castilho B. A., Olfson P., Casadaban M. J. Plasmid insertion mutagenesis and lac gene fusion with mini-mu bacteriophage transposons. J Bacteriol. 1984 May;158(2):488–495. doi: 10.1128/jb.158.2.488-495.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Chang A. C., Cohen S. N. Construction and characterization of amplifiable multicopy DNA cloning vehicles derived from the P15A cryptic miniplasmid. J Bacteriol. 1978 Jun;134(3):1141–1156. doi: 10.1128/jb.134.3.1141-1156.1978. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Chang J. Y., Knecht R., Braun D. G. Amino acid analysis at the picomole level. Application to the C-terminal sequence analysis of polypeptides. Biochem J. 1981 Dec 1;199(3):547–555. doi: 10.1042/bj1990547. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. Chu J., Shoeman R., Hart J., Coleman T., Mazaitis A., Kelker N., Brot N., Weissbach H. Cloning and expression of the metE gene in Escherichia coli. Arch Biochem Biophys. 1985 Jun;239(2):467–474. doi: 10.1016/0003-9861(85)90713-1. [DOI] [PubMed] [Google Scholar]
  11. Easton A. M., Kushner S. R. Transcription of the uvrD gene of Escherichia coli is controlled by the lexA repressor and by attenuation. Nucleic Acids Res. 1983 Dec 20;11(24):8625–8640. doi: 10.1093/nar/11.24.8625. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Finch P. W., Emmerson P. T. The nucleotide sequence of the uvrD gene of E. coli. Nucleic Acids Res. 1984 Jul 25;12(14):5789–5799. doi: 10.1093/nar/12.14.5789. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Girodeau J. M., Agouridas C., Masson M., Pineau R., Le Goffic F. The lysine pathway as a target for a new genera of synthetic antibacterial antibiotics? J Med Chem. 1986 Jun;29(6):1023–1030. doi: 10.1021/jm00156a021. [DOI] [PubMed] [Google Scholar]
  14. Hohn B., Collins J. A small cosmid for efficient cloning of large DNA fragments. Gene. 1980 Nov;11(3-4):291–298. doi: 10.1016/0378-1119(80)90069-4. [DOI] [PubMed] [Google Scholar]
  15. Homma H., Chiba N., Kobayashi T., Kudo I., Inoue K., Ikeda H., Sekiguchi M., Nojima S. Characteristics of detergent-resistant phospholipase A overproduced in E. coli cells bearing its cloned structural gene. J Biochem. 1984 Dec;96(6):1645–1653. doi: 10.1093/oxfordjournals.jbchem.a134996. [DOI] [PubMed] [Google Scholar]
  16. Koop A. H., Hartley M., Bourgeois S. Analysis of the cya locus of Escherichia coli. Gene. 1984 May;28(2):133–146. doi: 10.1016/0378-1119(84)90250-6. [DOI] [PubMed] [Google Scholar]
  17. Maples V. F., Kushner S. R. DNA repair in Escherichia coli: identification of the uvrD gene product. Proc Natl Acad Sci U S A. 1982 Sep;79(18):5616–5620. doi: 10.1073/pnas.79.18.5616. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Mechulam Y., Fromant M., Mellot P., Plateau P., Blanchin-Roland S., Fayat G., Blanquet S. Molecular cloning of the Escherichia coli gene for diadenosine 5',5'''-P1,P4-tetraphosphate pyrophosphohydrolase. J Bacteriol. 1985 Oct;164(1):63–69. doi: 10.1128/jb.164.1.63-69.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Mengin-Lecreulx D., Flouret B., van Heijenoort J. Cytoplasmic steps of peptidoglycan synthesis in Escherichia coli. J Bacteriol. 1982 Sep;151(3):1109–1117. doi: 10.1128/jb.151.3.1109-1117.1982. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Mengin-Lecreulx D., Flouret B., van Heijenoort J. Pool levels of UDP N-acetylglucosamine and UDP N-acetylglucosamine-enolpyruvate in Escherichia coli and correlation with peptidoglycan synthesis. J Bacteriol. 1983 Jun;154(3):1284–1290. doi: 10.1128/jb.154.3.1284-1290.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Misono H., Togawa H., Yamamoto T., Soda K. Meso-alpha,epsilon-diaminopimelate D-dehydrogenase: distribution and the reaction product. J Bacteriol. 1979 Jan;137(1):22–27. doi: 10.1128/jb.137.1.22-27.1979. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Monk M., Kinross J. Conditional lethality of recA and recB derivatives of a strain of Escherichia coli K-12 with a temperature-sensitive deoxyribonucleic acid polymerase I. J Bacteriol. 1972 Mar;109(3):971–978. doi: 10.1128/jb.109.3.971-978.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  23. Nilsson B., Uhlén M., Josephson S., Gatenbeck S., Philipson L. An improved positive selection plasmid vector constructed by oligonucleotide mediated mutagenesis. Nucleic Acids Res. 1983 Nov 25;11(22):8019–8030. doi: 10.1093/nar/11.22.8019. [DOI] [PMC free article] [PubMed] [Google Scholar]
  24. Roy A., Danchin A. Restriction map of the cya region of the Escherichia coli K12 chromosome. Biochimie. 1981 Aug-Sep;63(8-9):719–722. doi: 10.1016/s0300-9084(81)80220-9. [DOI] [PubMed] [Google Scholar]
  25. Thomas S. D., Jordan P. M. Nucleotide sequence of the hemC locus encoding porphobilinogen deaminase of Escherichia coli K12. Nucleic Acids Res. 1986 Aug 11;14(15):6215–6226. doi: 10.1093/nar/14.15.6215. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Winans S. C., Elledge S. J., Krueger J. H., Walker G. C. Site-directed insertion and deletion mutagenesis with cloned fragments in Escherichia coli. J Bacteriol. 1985 Mar;161(3):1219–1221. doi: 10.1128/jb.161.3.1219-1221.1985. [DOI] [PMC free article] [PubMed] [Google Scholar]
  27. Wiseman J. S., Nichols J. S. Purification and properties of diaminopimelic acid epimerase from Escherichia coli. J Biol Chem. 1984 Jul 25;259(14):8907–8914. [PubMed] [Google Scholar]
  28. Yanisch-Perron C., Vieira J., Messing J. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and pUC19 vectors. Gene. 1985;33(1):103–119. doi: 10.1016/0378-1119(85)90120-9. [DOI] [PubMed] [Google Scholar]
  29. de Geus P., van Die I., Bergmans H., Tommassen J., de Haas G. Molecular cloning of pldA, the structural gene for outer membrane phospholipase of E. coli K12. Mol Gen Genet. 1983;190(1):150–155. doi: 10.1007/BF00330338. [DOI] [PubMed] [Google Scholar]

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