Production and purification of Burkholderia pseudomallei BipD protein.
Type III secretion system (TTSS) clusters contain virulent genes of both animal and plant Gram-negative bacteria. The full length (933 bp) gene of bipD, a member of TTSS cluster of Burkholderia pseudomallei, was isolated by PCR amplification from B. pseudomallei DNA and cloned into pGEX 4T-1. GST-BipD fusion protein and BipD protein obtained by removal of GST using thrombin were employed to detect the presence of B. pseudomallei antibodies in sera obtained from melioidosis and non-melioidosis patients by immunoblotting. Sensitivity and specificity of BipD protein was 100% and 91.1%, respectively, whereas that of fusion protein was 77.8% and 90.0%, respectively.
Department of Biochemistry, Faculty of Science, Prince of Songkla University, Hat Yai, Songkhla, Thailand.
Monton
M
Visutthi
Siroj
S
Jitsurong
Wilaiwan
W
Chotigeat
D000595:Amino Acid Sequence; D000907:Antibodies, Bacterial / Q000097:blood; D001426:Bacterial Proteins / Q000235:genetics / Q000302:isolation & purification*; D016957:Burkholderia pseudomallei / Q000235:genetics / Q000276:immunology / Q000378:metabolism* / Q000472:pathogenicity; D016022:Case-Control Studies; D017931:DNA Primers; D050939:Gene Fusion; D005798:Genes, Bacterial; D006801:Humans; D008554:Melioidosis / Q000235:genetics; D008969:Molecular Sequence Data; D005810:Multigene Family; D016133:Polymerase Chain Reaction; D012680:Sensitivity and Specificity; D013785:Thailand; D014774:Virulence / Q000235:genetics
D000907:Antibodies, Bacterial; D001426:Bacterial Proteins; D017931:DNA Primers