Construction and characterization of transposon TnphoZ for the identification of genes encoding exported proteins in Streptococcus agalactiae.
Bacterial virulence often depends on exported proteins. To identify genes encoding exported proteins in the neonatal pathogen, group B streptococcus, the transposon TnphoZ was constructed. Here, the coding sequence for the secretion-dependent enzyme alkaline phosphatase from Enterococcus faecalis was fused to the left terminal repeat of Tn917, generating TnphoZ. A collection of TnphoZ mutants was isolated and the DNA flanking the transposon insertion sites was sequenced. Sequence data correlated the expression of high AP activity with transposon insertion into genes encoding predicted exported proteins. It is anticipated that TnphoZ will be suitable for use in other Gram-positive hosts.
Clancy
Anne
A
Department of Pediatrics, Division of Infectious Disease, Children's Hospital and Regional Medical Center, University of Washington, Seattle, WA 98105, USA. anne.clancy@seattlechildrens.org
Lee
Martin H
MH
Jones
Amanda L
AL
Rubens
Craig E
CE
D000469:Alkaline Phosphatase / Q000235:genetics / Q000378:metabolism; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D001483:Base Sequence; D004251:DNA Transposable Elements / Q000235:genetics*; D013293:Enterococcus faecalis / Q000201:enzymology / Q000235:genetics; D005821:Genetic Techniques; D006801:Humans; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D010957:Plasmids; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D017422:Sequence Analysis, DNA; D013292:Streptococcus agalactiae / Q000235:genetics / Q000378:metabolism*
D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D011993:Recombinant Fusion Proteins; D000469:Alkaline Phosphatase