Logo of jbacterPermissionsJournals.ASM.orgJournalJB ArticleJournal InfoAuthorsReviewers
J Bacteriol. 1990 Mar; 172(3): 1256–1261.
PMCID: PMC208591

Cloning, nucleotide sequence, and expression in Escherichia coli of the phospholipase D gene from Corynebacterium pseudotuberculosis.

Abstract

The phospholipase D (PLD) gene from Corynebacterium pseudotuberculosis has been cloned, sequenced, and expressed in Escherichia coli. Analysis of DNA sequence data reveals a major open reading frame encoding a 31.4-kilodalton protein, a size consistent with that estimated for the PLD protein by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Comparison of these data with the amino-terminal protein sequence indicates that the mature PLD protein is preceded by a 24-residue signal sequence. Expression of the PLD gene in E. coli is initiated from the corynebacterial promoter, and the resulting protein has sphingomyelinase activity. Primer extension mapping localized the 5' end of the PLD gene mRNA to a site 5 to 7 base pairs downstream of a region similar to the consensus sequence for E. coli promoters. Northern and Southern blot analyses suggest that the gene is transcribed from mRNA approximately 1.1 kilobases in length and that it is present in a single copy within the C. pseudotuberculosis genome.

Full text

Full text is available as a scanned copy of the original print version. Get a printable copy (PDF file) of the complete article (1.1M), or click on a page image below to browse page by page. Links to PubMed are also available for Selected References.

Images in this article

Click on the image to see a larger version.

Selected References

These references are in PubMed. This may not be the complete list of references from this article.
  • Aiba H, Adhya S, de Crombrugghe B. Evidence for two functional gal promoters in intact Escherichia coli cells. J Biol Chem. 1981 Nov 25;256(22):11905–11910. [PubMed]
  • Batey RG. Pathogenesis of caseous lymphadenitis in sheep and goats. Aust Vet J. 1986 Sep;63(9):269–272. [PubMed]
  • Boyd J, Murphy JR. Analysis of the diphtheria tox promoter by site-directed mutagenesis. J Bacteriol. 1988 Dec;170(12):5949–5952. [PMC free article] [PubMed]
  • Gatt S, Dinur T, Barenholz Y. A spectrophotometric method for determination of sphingomyelinase. Biochim Biophys Acta. 1978 Sep 28;530(3):503–507. [PubMed]
  • Greenfield L, Bjorn MJ, Horn G, Fong D, Buck GA, Collier RJ, Kaplan DA. Nucleotide sequence of the structural gene for diphtheria toxin carried by corynebacteriophage beta. Proc Natl Acad Sci U S A. 1983 Nov;80(22):6853–6857. [PMC free article] [PubMed]
  • Hsu TY, Renshaw HW, Livingston CW, Jr, Augustine JL, Zink DL, Gauer BB. Corynebacterium pseudotuberculosis exotoxin: fatal hemolytic anemia induced in gnotobiotic neonatal small ruminants by parenteral administration of preparations containing exotoxin. Am J Vet Res. 1985 May;46(5):1206–1211. [PubMed]
  • Kaczorek M, Delpeyroux F, Chenciner N, Streeck RE, Murphy JR, Boquet P, Tiollais P. Nucleotide sequence and expression of the diphtheria tox228 gene in Escherichia coli. Science. 1983 Aug 26;221(4613):855–858. [PubMed]
  • Kendall DA, Bock SC, Kaiser ET. Idealization of the hydrophobic segment of the alkaline phosphatase signal peptide. Nature. 1986 Jun 12;321(6071):706–708. [PubMed]
  • McClure WR. Mechanism and control of transcription initiation in prokaryotes. Annu Rev Biochem. 1985;54:171–204. [PubMed]
  • Meinkoth J, Wahl G. Hybridization of nucleic acids immobilized on solid supports. Anal Biochem. 1984 May 1;138(2):267–284. [PubMed]
  • Ohtsuka E, Matsuki S, Ikehara M, Takahashi Y, Matsubara K. An alternative approach to deoxyoligonucleotides as hybridization probes by insertion of deoxyinosine at ambiguous codon positions. J Biol Chem. 1985 Mar 10;260(5):2605–2608. [PubMed]
  • Soucková A, Soucek A. Inhibition of the hemolytic action of and lysins of Staphylococcus pyogenes by Corynebacterium hemolyticum, C. ovis and C. ulcerans. Toxicon. 1972 Aug;10(5):501–509. [PubMed]
  • Tabor S, Richardson CC. DNA sequence analysis with a modified bacteriophage T7 DNA polymerase. Proc Natl Acad Sci U S A. 1987 Jul;84(14):4767–4771. [PMC free article] [PubMed]
  • ZAKI MM. PRODUCTION OF A SOLUBLE SUBSTANCE BY CORYNEBACTERIUM OVIS. Nature. 1965 Feb 27;205:928–929. [PubMed]
  • Vieira J, Messing J. Production of single-stranded plasmid DNA. Methods Enzymol. 1987;153:3–11. [PubMed]
  • von Heijne G. Signal sequences. The limits of variation. J Mol Biol. 1985 Jul 5;184(1):99–105. [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

  • MedGen
    MedGen
    Related information in MedGen
  • Pathways + GO
    Pathways + GO
    Pathways, annotations and biological systems (BioSystems) that cite the current article.
  • Protein
    Protein
    Published protein sequences
  • PubMed
    PubMed
    PubMed citations for these articles
  • Substance
    Substance
    PubChem Substance links

Recent Activity

Your browsing activity is empty.

Activity recording is turned off.

Turn recording back on

See more...