Skip to main content
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1992 Dec;174(23):7784–7790. doi: 10.1128/jb.174.23.7784-7790.1992

Cloning of the Escherichia coli sor genes for L-sorbose transport and metabolism and physical mapping of the genes near metH and iclR.

U F Wehmeier 1, B Nobelmann 1, J W Lengeler 1
PMCID: PMC207494  PMID: 1447144

Abstract

The sor genes for L-sorbose (Sor) degradation of Escherichia coli EC3132, a wild-type strain, have been cloned on a 10.8-kbp fragment together with parts of the metH gene. The genes were mapped by restriction analysis, by deletion mapping, and by insertion mutagenesis with Tn1725. Seven sor genes with their corresponding gene products have been identified. They form an operon (gene order sorCpCDFBAME) inducible by L-sorbose, and their products have the following functions: SorC (36 kDa), regulatory protein with repressor-activator functions; SorD (29 kDa), D-glucitol-6-phosphate dehydrogenase; SorF and SorB (14 and 19 kDa, respectively), and SorA and SorM (27 and 29 kDa, respectively), two soluble and two membrane-bound proteins, respectively, of an L-sorbose phosphotransferase transport system; SorE (45 kDa), sorbose-1-phosphate reductase. The sor operon from E. coli EC3132 thus is identical to the operon from Klebsiella pneumoniae KAY2026. On the basis of restriction mapping followed by Southern hybridization experiments, the sor genes were mapped at 91.2 min on the chromosome, 3.3 kbp downstream of the metH-iclR gene cluster, and shown to be transcribed in a counterclockwise direction. The chromosomal map of the Sor+ strain EC3132 differs from that of the Sor- strain K-12 in approximately 8.6 kbp.

Full text

PDF
7784

Images in this article

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Altenbuchner J., Schmid K., Schmitt R. Tn1721-encoded tetracycline resistance: mapping of structural and regulatory genes mediating resistance. J Bacteriol. 1983 Jan;153(1):116–123. doi: 10.1128/jb.153.1.116-123.1983. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. 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: 10.1006/abio.1976.9999. [DOI] [PubMed] [Google Scholar]
  3. Brass J. M., Manson M. D. Reconstitution of maltose chemotaxis in Escherichia coli by addition of maltose-binding protein to calcium-treated cells of maltose regulon mutants. J Bacteriol. 1984 Mar;157(3):881–890. doi: 10.1128/jb.157.3.881-890.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  4. Brickman E., Soll L., Beckwith J. Genetic characterization of mutations which affect catabolite-sensitive operons in Escherichia coli, including deletions of the gene for adenyl cyclase. J Bacteriol. 1973 Nov;116(2):582–587. doi: 10.1128/jb.116.2.582-587.1973. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Fraenkel D. G., Levisohn S. R. Glucose and gluconate metabolism in an Escherichia coli mutant lacking phosphoglucose isomerase. J Bacteriol. 1967 May;93(5):1571–1578. doi: 10.1128/jb.93.5.1571-1578.1967. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Hill C. W., Grafstrom R. H., Harnish B. W., Hillman B. S. Tandem duplications resulting from recombination between ribosomal RNA genes in Escherichia coli. J Mol Biol. 1977 Nov 5;116(3):407–428. doi: 10.1016/0022-2836(77)90077-8. [DOI] [PubMed] [Google Scholar]
  7. Kelker N. E., Simkins R. A., Anderson R. L. Pathway of L-sorbose metabolism in Aerobacter aerogenes. J Biol Chem. 1972 Mar 10;247(5):1479–1483. [PubMed] [Google Scholar]
  8. Kohara Y., Akiyama K., Isono K. The physical map of the whole E. coli chromosome: application of a new strategy for rapid analysis and sorting of a large genomic library. Cell. 1987 Jul 31;50(3):495–508. doi: 10.1016/0092-8674(87)90503-4. [DOI] [PubMed] [Google Scholar]
  9. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  10. Lengeler J., Lin E. C. Reversal of the mannitol-sorbitol diauxie in Escherichia coli. J Bacteriol. 1972 Nov;112(2):840–848. doi: 10.1128/jb.112.2.840-848.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lengeler J. Mutations affecting transport of the hexitols D-mannitol, D-glucitol, and galactitol in Escherichia coli K-12: isolation and mapping. J Bacteriol. 1975 Oct;124(1):26–38. doi: 10.1128/jb.124.1.26-38.1975. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Médigue C., Bouché J. P., Hénaut A., Danchin A. Mapping of sequenced genes (700 kbp) in the restriction map of the Escherichia coli chromosome. Mol Microbiol. 1990 Feb;4(2):169–187. doi: 10.1111/j.1365-2958.1990.tb00585.x. [DOI] [PubMed] [Google Scholar]
  13. Nègre D., Cortay J. C., Old I. G., Galinier A., Richaud C., Saint Girons I., Cozzone A. J. Overproduction and characterization of the iclR gene product of Escherichia coli K-12 and comparison with that of Salmonella typhimurium LT2. Gene. 1991 Jan 2;97(1):29–37. doi: 10.1016/0378-1119(91)90006-w. [DOI] [PubMed] [Google Scholar]
  14. Old I. G., Margarita D., Glass R. E., Saint Girons I. Nucleotide sequence of the metH gene of Escherichia coli K-12 and comparison with that of Salmonella typhimurium LT2. Gene. 1990 Mar 1;87(1):15–21. doi: 10.1016/0378-1119(90)90490-i. [DOI] [PubMed] [Google Scholar]
  15. Riley M., Anilionis A. Evolution of the bacterial genome. Annu Rev Microbiol. 1978;32:519–560. doi: 10.1146/annurev.mi.32.100178.002511. [DOI] [PubMed] [Google Scholar]
  16. Short J. M., Fernandez J. M., Sorge J. A., Huse W. D. Lambda ZAP: a bacteriophage lambda expression vector with in vivo excision properties. Nucleic Acids Res. 1988 Aug 11;16(15):7583–7600. doi: 10.1093/nar/16.15.7583. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Sprenger G. A., Lengeler J. W. L-Sorbose metabolism in Klebsiella pneumoniae and Sor+ derivatives of Escherichia coli K-12 and chemotaxis toward sorbose. J Bacteriol. 1984 Jan;157(1):39–45. doi: 10.1128/jb.157.1.39-45.1984. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Tabor S., Richardson C. C. A bacteriophage T7 RNA polymerase/promoter system for controlled exclusive expression of specific genes. Proc Natl Acad Sci U S A. 1985 Feb;82(4):1074–1078. doi: 10.1073/pnas.82.4.1074. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Van Gijsegem F., Toussaint A. Chromosome transfer and R-prime formation by an RP4::mini-Mu derivative in Escherichia coli, Salmonella typhimurium, Klebsiella pneumoniae, and Proteus mirabilis. Plasmid. 1982 Jan;7(1):30–44. doi: 10.1016/0147-619x(82)90024-5. [DOI] [PubMed] [Google Scholar]
  20. Wehmeier U., Sprenger G. A., Lengeler J. W. The use of lambda plac-Mu hybrid phages in Klebsiella pneumoniae and the isolation of stable Hfr strains. Mol Gen Genet. 1989 Feb;215(3):529–536. doi: 10.1007/BF00427052. [DOI] [PubMed] [Google Scholar]
  21. Woodward M. J., Charles H. P. Genes for l-sorbose utilization in Escherichia coli. J Gen Microbiol. 1982 Sep;128(9):1969–1980. doi: 10.1099/00221287-128-9-1969. [DOI] [PubMed] [Google Scholar]
  22. Wöhrl B. M., Lengeler J. W. Cloning and physical mapping of the sor genes for L-sorbose transport and metabolism from Klebsiella pneumoniae. Mol Microbiol. 1990 Sep;4(9):1557–1565. doi: 10.1111/j.1365-2958.1990.tb02067.x. [DOI] [PubMed] [Google Scholar]
  23. Wöhrl B. M., Sprenger G. A., Lengeler J. W. Construction of a new catabolic pathway for D-fructose in Escherichia coli K12 using an L-sorbose-specific enzyme from Klebsiella pneumoniae. Arch Microbiol. 1990;154(2):162–167. doi: 10.1007/BF00423327. [DOI] [PubMed] [Google Scholar]
  24. Wöhrl B. M., Wehmeier U. F., Lengeler J. W. Positive and negative regulation of expression of the L-sorbose (sor) operon by SorC in Klebsiella pneumoniae. Mol Gen Genet. 1990 Nov;224(2):193–200. doi: 10.1007/BF00271552. [DOI] [PubMed] [Google Scholar]
  25. 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]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES