• We are sorry, but NCBI web applications do not support your browser and may not function properly. More information
Logo of jbacterPermissionsJournals.ASM.orgJournalJB ArticleJournal InfoAuthorsReviewers
J Bacteriol. Jun 1996; 178(12): 3614–3620.
PMCID: PMC178134

Polar allele duplication for transcriptional analysis of consecutive essential genes: application to a cluster of Escherichia coli fatty acid biosynthetic genes.

Abstract

The genes encoding acyl carrier protein and several key fatty acid biosynthetic enzymes are clustered at min 24 of the Escherichia coli chromosome. This cluster of genes is not transcribed as a classical operon, but rather multiple promoters are present and each gene is cotranscribed with at least one other gene. Transcripts specific for single genes ar also present. Transcription of acpP, the gene encoding acyl carrier protein, has been studied in detail. The acpP gene was shown to be transcribed from at least two different promoters by Northern (RNA) blot, primer extension, and deletion analyses, and the major promoter was mapped. We tested whether multiple promoters are necessary to produce acyl carrier protein by use of a new method whereby a transcriptional terminator was integrated into the chromosome upstream of the intact acpP gene. By use of this method (called polar allele duplication), we demonstrate that the promoter located immediately upstream of the coding sequence is sufficient for synthesis of this very abundant protein.

Full Text

The Full Text of this article is available as a PDF (580K).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Aldea M, Garrido T, Pla J, Vicente M. Division genes in Escherichia coli are expressed coordinately to cell septum requirements by gearbox promoters. EMBO J. 1990 Nov;9(11):3787–3794. [PMC free article] [PubMed]
  • Arps PJ, Winkler ME. Structural analysis of the Escherichia coli K-12 hisT operon by using a kanamycin resistance cassette. J Bacteriol. 1987 Mar;169(3):1061–1070. [PMC free article] [PubMed]
  • Chang YY, Cronan JE., Jr Mapping nonselectable genes of Escherichia coli by using transposon Tn10: location of a gene affecting pyruvate oxidase. J Bacteriol. 1982 Sep;151(3):1279–1289. [PMC free article] [PubMed]
  • Fellay R, Frey J, Krisch H. Interposon mutagenesis of soil and water bacteria: a family of DNA fragments designed for in vitro insertional mutagenesis of gram-negative bacteria. Gene. 1987;52(2-3):147–154. [PubMed]
  • Garwin JL, Klages AL, Cronan JE., Jr Structural, enzymatic, and genetic studies of beta-ketoacyl-acyl carrier protein synthases I and II of Escherichia coli. J Biol Chem. 1980 Dec 25;255(24):11949–11956. [PubMed]
  • Heath RJ, Rock CO. Enoyl-acyl carrier protein reductase (fabI) plays a determinant role in completing cycles of fatty acid elongation in Escherichia coli. J Biol Chem. 1995 Nov 3;270(44):26538–26542. [PubMed]
  • Henry MF, Cronan JE., Jr A new mechanism of transcriptional regulation: release of an activator triggered by small molecule binding. Cell. 1992 Aug 21;70(4):671–679. [PubMed]
  • Jackowski S, Rock CO. Altered molecular form of acyl carrier protein associated with beta-ketoacyl-acyl carrier protein synthase II (fabF) mutants. J Bacteriol. 1987 Apr;169(4):1469–1473. [PMC free article] [PubMed]
  • Kornblum JS, Projan SJ, Moghazeh SL, Novick RP. A rapid method to quantitate non-labeled RNA species in bacterial cells. Gene. 1988;63(1):75–85. [PubMed]
  • Larson TJ, Ludtke DN, Bell RM. sn-Glycerol-3-phosphate auxotrophy of plsB strains of Escherichia coli: evidence that a second mutation, plsX, is required. J Bacteriol. 1984 Nov;160(2):711–717. [PMC free article] [PubMed]
  • Lee SY, Rasheed S. A simple procedure for maximum yield of high-quality plasmid DNA. Biotechniques. 1990 Dec;9(6):676–679. [PubMed]
  • Li SJ, Cronan JE., Jr Growth rate regulation of Escherichia coli acetyl coenzyme A carboxylase, which catalyzes the first committed step of lipid biosynthesis. J Bacteriol. 1993 Jan;175(2):332–340. [PMC free article] [PubMed]
  • Magnuson K, Carey MR, Cronan JE., Jr The putative fabJ gene of Escherichia coli fatty acid synthesis is the fabF gene. J Bacteriol. 1995 Jun;177(12):3593–3595. [PMC free article] [PubMed]
  • Magnuson K, Jackowski S, Rock CO, Cronan JE., Jr Regulation of fatty acid biosynthesis in Escherichia coli. Microbiol Rev. 1993 Sep;57(3):522–542. [PMC free article] [PubMed]
  • Magnuson K, Oh W, Larson TJ, Cronan JE., Jr Cloning and nucleotide sequence of the fabD gene encoding malonyl coenzyme A-acyl carrier protein transacylase of Escherichia coli. FEBS Lett. 1992 Mar 16;299(3):262–266. [PubMed]
  • Metcalf WW, Jiang W, Wanner BL. Use of the rep technique for allele replacement to construct new Escherichia coli hosts for maintenance of R6K gamma origin plasmids at different copy numbers. Gene. 1994 Jan 28;138(1-2):1–7. [PubMed]
  • Mulligan ME, Hawley DK, Entriken R, McClure WR. Escherichia coli promoter sequences predict in vitro RNA polymerase selectivity. Nucleic Acids Res. 1984 Jan 11;12(1 Pt 2):789–800. [PMC free article] [PubMed]
  • Oh W, Larson TJ. Physical locations of genes in the rne (ams)-rpmF-plsX-fab region of the Escherichia coli K-12 chromosome. J Bacteriol. 1992 Dec;174(23):7873–7874. [PMC free article] [PubMed]
  • Ohlrogge JB, Browse J, Somerville CR. The genetics of plant lipids. Biochim Biophys Acta. 1991 Feb 26;1082(1):1–26. [PubMed]
  • Podkovyrov S, Larson TJ. Lipid biosynthetic genes and a ribosomal protein gene are cotranscribed. FEBS Lett. 1995 Jul 24;368(3):429–431. [PubMed]
  • Post-Beittenmiller D, Jaworski JG, Ohlrogge JB. In vivo pools of free and acylated acyl carrier proteins in spinach. Evidence for sites of regulation of fatty acid biosynthesis. J Biol Chem. 1991 Jan 25;266(3):1858–1865. [PubMed]
  • Rawlings M, Cronan JE., Jr The gene encoding Escherichia coli acyl carrier protein lies within a cluster of fatty acid biosynthetic genes. J Biol Chem. 1992 Mar 25;267(9):5751–5754. [PubMed]
  • Takeshita S, Sato M, Toba M, Masahashi W, Hashimoto-Gotoh T. High-copy-number and low-copy-number plasmid vectors for lacZ alpha-complementation and chloramphenicol- or kanamycin-resistance selection. Gene. 1987;61(1):63–74. [PubMed]
  • Tsay JT, Oh W, Larson TJ, Jackowski S, Rock CO. Isolation and characterization of the beta-ketoacyl-acyl carrier protein synthase III gene (fabH) from Escherichia coli K-12. J Biol Chem. 1992 Apr 5;267(10):6807–6814. [PubMed]

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

Formats:

Related citations in PubMed

See reviews...See all...

Cited by other articles in PMC

See all...

Links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...