Alkaline phosphatase reporter transposon for identification of genes encoding secreted proteins in gram-positive microorganisms.
We describe the construction of TnFuZ, a genetic tool for the discovery and mutagenesis of proteins exported from gram-positive bacteria. This tool combines a transposable element (Tn4001) of broad host range in gram-positive bacteria and an alkaline phosphatase gene (phoZ) derived from a gram-positive bacterium that has been modified by removal of the region encoding its export signal. Mutagenesis of Streptococcus pyogenes with TnFuZ ("FuZ" stands for fusions to phoZ) identified genes encoding secreted proteins whose expression was enhanced during growth in an aerobic environment. Thus, TnFuZ should be valuable for analysis of protein secretion, gene regulation, and virulence in gram-positive bacteria.
Gibson
Carmela M
CM
Department of Molecular Microbiology, University Medical Center, Washington University School of Medicine, 660 S. Euclid Ave., St. Louis, Missouri 63110-1093, USA.
Caparon
Michael G
MG
D000469:Alkaline Phosphatase / Q000235:genetics*; D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D001483:Base Sequence; D004251:DNA Transposable Elements / Q000235:genetics*; D018274:Electroporation; D015964:Gene Expression Regulation, Bacterial; D017930:Genes, Reporter; D006094:Gram-Positive Bacteria / Q000235:genetics* / Q000378:metabolism / Q000472:pathogenicity; D006801:Humans; D008969:Molecular Sequence Data; D016254:Mutagenesis, Insertional; D017422:Sequence Analysis, DNA; D013297:Streptococcus pyogenes / Q000235:genetics / Q000378:metabolism / Q000472:pathogenicity; D014169:Transformation, Bacterial
D001426:Bacterial Proteins; D004251:DNA Transposable Elements; D000469:Alkaline Phosphatase