Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1981 Sep;147(3):797–804. doi: 10.1128/jb.147.3.797-804.1981

Physical characterization of the ilvHI operon of Escherichia coli K-12.

C H Squires, M De Felice, S R Wessler, J M Calvo
PMCID: PMC216115  PMID: 6168634

Abstract

The ilvHI and leu genes of Escherichia coli K-12 are contained on a single 10.9-kilobase EcoRI fragment of deoxyribonucleic acid derived from the leu transducing phage lambda G4. Since the expression of all of these genes is controlled by leucine, we investigated whether they are part of single operon or whether they constitute separate but adjacent operons controlled from a common site. Both cloning and hybridization studies indicated that ilvHI and leu are distinct operons. They are transcribed in opposite directions and are separated by approximately 1,500 base pairs of deoxyribonucleic acid. Hybridization experiments showed that the expression of ilvHI is regulated chiefly at the level of transcription. The size of the ilvHI messenger ribonucleic acid is estimated to be 2,550 bases.

Full text

PDF
801

Images in this article

802Image
on p.802

Selected References

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

  1. Alwine J. C., Kemp D. J., Stark G. R. Method for detection of specific RNAs in agarose gels by transfer to diazobenzyloxymethyl-paper and hybridization with DNA probes. Proc Natl Acad Sci U S A. 1977 Dec;74(12):5350–5354. doi: 10.1073/pnas.74.12.5350. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bachmann B. J., Low K. B. Linkage map of Escherichia coli K-12, edition 6. Microbiol Rev. 1980 Mar;44(1):1–56. doi: 10.1128/mr.44.1.1-56.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bailey J. M., Davidson N. Methylmercury as a reversible denaturing agent for agarose gel electrophoresis. Anal Biochem. 1976 Jan;70(1):75–85. doi: 10.1016/s0003-2697(76)80049-8. [DOI] [PubMed] [Google Scholar]
  4. Berg C. M., Shaw K. J., Berg D. E. The ilvG gene is expressed in Escherichia coli K-12. Gene. 1980 Dec;12(1-2):165–170. doi: 10.1016/0378-1119(80)90028-1. [DOI] [PubMed] [Google Scholar]
  5. Calvo J. M., Freundlich M., Umbarger H. E. Regulation of branched-chain amino acid biosynthesis in Salmonella typhimurium: isolation of regulatory mutants. J Bacteriol. 1969 Mar;97(3):1272–1282. doi: 10.1128/jb.97.3.1272-1282.1969. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Cooper T. G., Whitney P., Magasanik B. Reaction of lac-specific ribonucleic acid from Escherichia coli with lac deoxyribonucleic acid. J Biol Chem. 1974 Oct 25;249(20):6548–6555. [PubMed] [Google Scholar]
  7. Davis M. G., Calvo J. M. Isolation and characterization of lambda pleu bacteriophages. J Bacteriol. 1977 Feb;129(2):1078–1090. doi: 10.1128/jb.129.2.1078-1090.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Davis M. G., Calvo J. M. Relationship between messenger ribonucleic acid and enzyme levels specified by the leucine operon of Escherichia coli K-12. J Bacteriol. 1977 Sep;131(3):997–1007. doi: 10.1128/jb.131.3.997-1007.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. De Felice M., Guardiola J., Esposito B., Iaccarino M. Structural genes for a newly recognized acetolactate synthase in Escherichia coli K-12. J Bacteriol. 1974 Dec;120(3):1068–1077. doi: 10.1128/jb.120.3.1068-1077.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  10. De Felice M., Guardiola J., Malorni M. C., Klopotowski T., Iaccarino M. Regulation of the pool size of valine in Escherichia coli K-12. J Bacteriol. 1974 Dec;120(3):1058–1067. doi: 10.1128/jb.120.3.1058-1067.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. De Felice M., Levinthal M. The acetohydroxy acid synthase III isoenzyme of Escherichia coli K-12: regulation of synthesis by leucine. Biochem Biophys Res Commun. 1977 Nov 7;79(1):82–87. doi: 10.1016/0006-291x(77)90063-8. [DOI] [PubMed] [Google Scholar]
  12. De Felice M., Squires C., Levinthal M., Guardiola J., Lamberti A., Iaccarino M. Growth inhibition of Escherichia coli K-12 by L-valine: a consequence of a regulatory pattern. Mol Gen Genet. 1977 Nov 4;156(1):1–7. doi: 10.1007/BF00272245. [DOI] [PubMed] [Google Scholar]
  13. Favre R., Wiater A., Puppo S., Iaccarino M. Expression of a valine-resistant acetolactate synthase activity mediated by the ilv O and ilv G genes of Escherichia coli K-12. Mol Gen Genet. 1976 Feb 2;143(3):243–252. doi: 10.1007/BF00269400. [DOI] [PubMed] [Google Scholar]
  14. Groves W. E., Davis F. C., Jr, Sells B. H. Spectrophotometric determination of microgram quantities of protein without nucleic acid interference. Anal Biochem. 1968 Feb;22(2):195–210. doi: 10.1016/0003-2697(68)90307-2. [DOI] [PubMed] [Google Scholar]
  15. Guardiola J., De Felice M., Iaccarino M. Mutant of Escherichia coli K-12 missing acetolactate synthase activity. J Bacteriol. 1974 Oct;120(1):536–538. doi: 10.1128/jb.120.1.536-538.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Guardiola J., De Felice M., Lamberti A., Iaccarino M. The acetolactate synthase isoenzymes of Escherichia coli K-12. Mol Gen Genet. 1977 Nov 4;156(1):17–25. doi: 10.1007/BF00272247. [DOI] [PubMed] [Google Scholar]
  17. Lawther R. P., Calhoun D. H., Adams C. W., Hauser C. A., Gray J., Hatfield G. W. Molecular basis of valine resistance in Escherichia coli K-12. Proc Natl Acad Sci U S A. 1981 Feb;78(2):922–925. doi: 10.1073/pnas.78.2.922. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Rigby P. W., Dieckmann M., Rhodes C., Berg P. Labeling deoxyribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. J Mol Biol. 1977 Jun 15;113(1):237–251. doi: 10.1016/0022-2836(77)90052-3. [DOI] [PubMed] [Google Scholar]
  19. Smith D. R., Calvo J. M. Nucleotide sequence of the E coli gene coding for dihydrofolate reductase. Nucleic Acids Res. 1980 May 24;8(10):2255–2274. doi: 10.1093/nar/8.10.2255. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. Smith D. R., Calvo J. M. Regulation of dihydrofolate reductase synthesis in Escherichia coli. Mol Gen Genet. 1979 Aug;175(1):31–38. doi: 10.1007/BF00267852. [DOI] [PubMed] [Google Scholar]
  21. Subrahmanyam C. S., McCorkle G. M., Umbarger H. E. Physical location of the ilvO determinant in Escherichia coli K-12 deoxyribonucleic acid. J Bacteriol. 1980 May;142(2):547–555. doi: 10.1128/jb.142.2.547-555.1980. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Wahl G. M., Stern M., Stark G. R. Efficient transfer of large DNA fragments from agarose gels to diazobenzyloxymethyl-paper and rapid hybridization by using dextran sulfate. Proc Natl Acad Sci U S A. 1979 Aug;76(8):3683–3687. doi: 10.1073/pnas.76.8.3683. [DOI] [PMC free article] [PubMed] [Google Scholar]

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

RESOURCES