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. 1978 Jan;75(1):89–93. doi: 10.1073/pnas.75.1.89

Physical organization of the ilvEDAC genes of Escherichia coli strain K-12

George M McCorkle 1, Timothy D Leathers 1, H E Umbarger 1
PMCID: PMC411189  PMID: 343113

Abstract

We have determined the physical location of the ilvEDAC genes on the restriction cleavage map of the ilv region of Escherichia coli K-12 by two methods: (i) heteroduplex and endonuclease cleavage analysis of hybrid phages carrying genetically defined parts of the ilv cluster and (ii) complementation analysis and enzyme assays to determine ilv gene expression from hybrid plasmids containing DNA restriction fragments of the transducing phage λh80dilv. The ilvEDA and ilvC operons occupy 2.4 and 0.9 megadalton sequences of DNA, respectively, and are separated by a region of 0.6-0.75 megadalton. The ilvD region, specifying dihydroxy acid dehydrase, has a maximum coding capacity of about 55,000 daltons of polypeptide. Our results confirm that ilvC is transcribed clockwise on the E. coli K-12 map, in the same direction as ilaEDA. A secondary λ attachment site within ilvC has been located on a small (0.45 megadalton) EcoRI fragment. Our results are compared to other physical studies of ilv DNA.

Keywords: isoleucine and valine biosynthesis, restriction endonuclease mapping, heteroduplex analysis, gene expression

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Selected References

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  1. Avitabile A., Carlomagno-Cerillo S., Favvre R., Blasi F. Isolation of transducing bacteriophages for the histidine and isoleucine-valine operons in Escherichia coli K-12. J Bacteriol. 1972 Oct;112(1):40–47. doi: 10.1128/jb.112.1.40-47.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Bachmann B. J., Low K. B., Taylor A. L. Recalibrated linkage map of Escherichia coli K-12. Bacteriol Rev. 1976 Mar;40(1):116–167. doi: 10.1128/br.40.1.116-167.1976. [DOI] [PMC free article] [PubMed] [Google Scholar]
  3. Bolivar F., Rodriguez R. L., Betlach M. C., Boyer H. W. Construction and characterization of new cloning vehicles. I. Ampicillin-resistant derivatives of the plasmid pMB9. Gene. 1977;2(2):75–93. doi: 10.1016/0378-1119(77)90074-9. [DOI] [PubMed] [Google Scholar]
  4. Bolivar F., Rodriguez R. L., Greene P. J., Betlach M. C., Heyneker H. L., Boyer H. W., Crosa J. H., Falkow S. Construction and characterization of new cloning vehicles. II. A multipurpose cloning system. Gene. 1977;2(2):95–113. [PubMed] [Google Scholar]
  5. Calhoun D. H., Rimerman R. A., Hatfield G. W. Threonine deaminase from Escherichia coli. I. Purification and properties. J Biol Chem. 1973 May 25;248(10):3511–3516. [PubMed] [Google Scholar]
  6. Cohen B. M., Jones E. W. New Map Location of ilvO in ESCHERICHIA COLI. Genetics. 1976 Jun;83(2):201–225. doi: 10.1093/genetics/83.2.201. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. 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]
  8. Hershfield V., Boyer H. W., Yanofsky C., Lovett M. A., Helinski D. R. Plasmid ColEl as a molecular vehicle for cloning and amplification of DNA. Proc Natl Acad Sci U S A. 1974 Sep;71(9):3455–3459. doi: 10.1073/pnas.71.9.3455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  9. Hofler J. G., Decedue C. J., Luginbuhl G. H., Reynolds J. A., Burns R. O. The subunit structure of alpha-acetohydroxyacid isomeroreductase from Salmonella typhimurium. J Biol Chem. 1975 Feb 10;250(3):877–882. [PubMed] [Google Scholar]
  10. Kenerley M. E., Morgan E. A., Post L., Lindahl L., Nomura M. Characterization of hybrid plasmids carrying individual ribosomal ribonucleic acid transcription units of Escherichia coli. J Bacteriol. 1977 Dec;132(3):931–949. doi: 10.1128/jb.132.3.931-949.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Kline E. L., Brown C. S., Coleman W. G., Jr, Umbarger H. E. Regulation of isoleucine-valine biosynthesis in an ilvDAC deletion strain of Escherichia coli K-12. Biochem Biophys Res Commun. 1974 Apr 23;57(4):1144–1151. doi: 10.1016/0006-291x(74)90816-x. [DOI] [PubMed] [Google Scholar]
  12. Lederberg E. M., Cohen S. N. Transformation of Salmonella typhimurium by plasmid deoxyribonucleic acid. J Bacteriol. 1974 Sep;119(3):1072–1074. doi: 10.1128/jb.119.3.1072-1074.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Lee H. J., Otsubo E., Deonier R. C., Davidson N. Electron microscope heteroduplex studies of sequence relations among plasmids of Escherichia coli. V. ilv+ Deletion mutants of F14. J Mol Biol. 1974 Nov 15;89(4):585–597. doi: 10.1016/0022-2836(74)90037-0. [DOI] [PubMed] [Google Scholar]
  14. Marsh N. J., Duggan D. E. Ordering of mutant sites in the isoleucine-valine genes of Escherichia coli by use of merogenotes derived from F 14 : a new procedure for fine-structure mapping. J Bacteriol. 1972 Feb;109(2):730–740. doi: 10.1128/jb.109.2.730-740.1972. [DOI] [PMC free article] [PubMed] [Google Scholar]
  15. Meagher R. B., Shepherd R. J., Boyer H. W. The structure of cauliflower mosaic virus. I. A restriction endonuclease map of cauliflower mosaic virus DNA. Virology. 1977 Jul 15;80(2):362–375. doi: 10.1016/s0042-6822(77)80012-3. [DOI] [PubMed] [Google Scholar]
  16. Monnier N., Montmitonnet A., Chesne S., Pelmont J. Transaminase B d'Escherichia coli. I. - Purification et premières propriétés. Biochimie. 1976;58(6):663–675. doi: 10.1016/s0300-9084(76)80390-2. [DOI] [PubMed] [Google Scholar]
  17. Morgan E. A., Ikemura T., Nomura M. Identification of spacer tRNA genes in individual ribosomal RNA transcription units of Escherichia coli. Proc Natl Acad Sci U S A. 1977 Jul;74(7):2710–2714. doi: 10.1073/pnas.74.7.2710. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Otsubo E., Soll L., Deonier R. C., Lee H. J., Davidson N. Electron microscope heteroduplex studies of sequence relations among plasmids of Escherichia coli. VIII. The structure of bacteriophage phi 80d-3ilv+su+7, including the mapping of the ribosomal RNA genes. J Mol Biol. 1974 Nov 15;89(4):631–646. doi: 10.1016/0022-2836(74)90040-0. [DOI] [PubMed] [Google Scholar]
  19. RAMAKRISHNAN T., ADELBERG E. A. REGULATORY MECHANISMS IN THE BIOSYNTHESIS OF ISOLEUCINE AND VALINE. 3. MAP ORDER OF THE STRUCTURAL GENES AND OPERATOR GENES. J Bacteriol. 1965 Mar;89:661–664. doi: 10.1128/jb.89.3.661-664.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  20. RAMAKRISHNAN T., ADELBERG E. A. REGULATORY MECHANISMS IN THE BIOSYNTHESIS OF ISOLEUCINE AND VALINE. II. IDENTIFICATION OF TWO OPERATOR GENES. J Bacteriol. 1965 Mar;89:654–660. doi: 10.1128/jb.89.3.654-660.1965. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Ratzkin B., Carbon J. Functional expression of cloned yeast DNA in Escherichia coli. Proc Natl Acad Sci U S A. 1977 Feb;74(2):487–491. doi: 10.1073/pnas.74.2.487. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Schiavo F. L., Favre R., Kasai T., Cascino A., Guardiola J., Caro L., Iaccarino M. The phi80 lambdadilv phage and its use to detect messenger RNA related to isoleucine-valine metabolism. J Mol Biol. 1975 Dec 15;99(3):353–368. doi: 10.1016/s0022-2836(75)80131-8. [DOI] [PubMed] [Google Scholar]
  23. Smith J. M., Smolin D. E., Umbarger H. E. Polarity and the regulation of the ilv gene cluster in Escherichia coli strain K-12. Mol Gen Genet. 1976 Oct 18;148(2):111–124. doi: 10.1007/BF00268374. [DOI] [PubMed] [Google Scholar]
  24. Smith J. M., Umbarger H. E. Characterization of fusions between the lac operon and the ilv gene cluster in Escherichia coli: ilvC-lac fusions. J Bacteriol. 1977 Dec;132(3):870–875. doi: 10.1128/jb.132.3.870-875.1977. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Vonder Haar R. A., Umbarger H. E. Isoleucine and valine metabolism in Escherichia coli K-12: detection and measurement of ilv-specific messenger ribonucleic acid. J Bacteriol. 1974 Nov;120(2):687–696. doi: 10.1128/jb.120.2.687-696.1974. [DOI] [PMC free article] [PubMed] [Google Scholar]
  26. Westmoreland B. C., Szybalski W., Ris H. Mapping of deletions and substitutions in heteroduplex DNA molecules of bacteriophage lambda by electron microscopy. Science. 1969 Mar 21;163(3873):1343–1348. doi: 10.1126/science.163.3873.1343. [DOI] [PubMed] [Google Scholar]

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