Cloning and characterization of a nonhemolytic phospholipase C gene from Burkholderia pseudomallei

J Clin Microbiol. 1999 Nov;37(11):3742-5. doi: 10.1128/JCM.37.11.3742-3745.1999.

Abstract

We cloned and characterized a phosphatidylcholine-hydrolyzing phospholipase C (PC-PLC) gene from Burkholderia pseudomallei. DNA sequence analysis of the gene indicated an open reading frame coding for 700 amino acids with a 34-amino-acid signal peptide. When cleaved, this yields a secreted 73-kDa mature protein. The deduced amino acid sequence exhibited 48% similarity to that of a nonhemolytic PLC from Pseudomonas aeruginosa. The expressed PC-PLC was heat stable, nonhemolytic for sheep erythrocytes, and active between pH 2 and 8. Western blot analysis with sera from melioidosis patients indicated that they produced immunoglobulin M antibodies against this PC-PLC protein.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antibodies, Bacterial / blood
  • Base Sequence
  • Burkholderia pseudomallei / enzymology*
  • Burkholderia pseudomallei / genetics*
  • Burkholderia pseudomallei / immunology
  • Cloning, Molecular
  • DNA Primers / genetics
  • DNA, Bacterial / genetics
  • Genes, Bacterial*
  • Hemolysis
  • Humans
  • Immunoglobulin M / blood
  • In Vitro Techniques
  • Melioidosis / immunology
  • Melioidosis / microbiology
  • Molecular Sequence Data
  • Restriction Mapping
  • Sheep
  • Type C Phospholipases / genetics*

Substances

  • Antibodies, Bacterial
  • DNA Primers
  • DNA, Bacterial
  • Immunoglobulin M
  • Type C Phospholipases
  • phosphatidylcholine-specific phospholipase C

Associated data

  • GENBANK/AF107252