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Cloning and characterization of a nonhemolytic phospholipase C gene from Burkholderia pseudomallei
S Korbsrisate1, N Suwanasai, A Leelaporn
1Departments of Immunology, Faculty of Medicine Siriraj Hospital, Bangkok 10700, Thailand. grsks@mahidol.ac.th
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.