title: Searching for virulence Burkholderia pseudomallei genes by immunoscreening the lambda ZAPII expressed genomic library.
abstract: The lambdaZAP II expressed genomic library of B. pseudomallei was screened with pooled melioidosis serum preabsorbed with E. coli host cell. The positive clones were detected by using protein A-CDP-star chemiluminescence. All of 14 positive clones reacted with only the pooled absorbed melioidosis serum and not the pooled absorbed normal serum when tested with the plaque dot blot analysis. The expressed genes were detected by using a combination of immunoscreening, bioinformatics and molecular biology. At least six in vivo expressed genes were identified by this approach. Two were well known virulent genes, gmhA (a capsule biosynthetic gene) and bipD (type III secretion protein gene). Another two were genes coded for conserved hypothetical protein. The last two isolated genes were groEL (a chaperonine protein gene), and a gene encoding transmembrane protein.
authors.affiliation: Inter-Department of Medical Microbiology, Graduate School and Prince of Songkla University, Songkhla, Thailand.
authors.forename: Siroj
authors.initials: S
authors.lastname: Jitsurong
authors.forename: Chintana
authors.initials: C
authors.lastname: Chirathaworn
authors.forename: Nat F
authors.initials: NF
authors.lastname: Brown
authors.forename: Ifor R
authors.initials: IR
authors.lastname: Beacham
authors.forename: Somying
authors.initials: S
authors.lastname: Tumwasorn
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics*; D010582:Bacteriophage lambda; D016957:Burkholderia pseudomallei / Q000235:genetics* / Q000472:pathogenicity*; D003001:Cloning, Molecular; D020869:Gene Expression Profiling; D015698:Genomic Library; D006801:Humans; D007118:Immunoassay; D008163:Luminescent Measurements; D008554:Melioidosis / Q000382:microbiology*; D014774:Virulence / Q000235:genetics
chemical_list: D001426:Bacterial Proteins