TnphoA: a transposon probe for protein export signals.
We constructed a derivative of transposon Tn5 that permits the generation of hybrid proteins composed of alkaline phosphatase (EC 3.1.3.1) lacking its signal peptide fused to amino-terminal sequences of other proteins. Such a hybrid gives alkaline phosphatase activity if the protein fused to alkaline phosphatase contributes sequences that promote export and thus compensate for the missing alkaline phosphatase signal peptide. Fusions to both a secreted periplasmic protein and a complex cytoplasmic membrane protein led to alkaline phosphatase activity. TnphoA fusions should help localize export signals within the structure of a protein, such as a transmembrane protein, as well as identify new chromosomal genes for secreted and transmembrane proteins.
D000469:Alkaline Phosphatase / Q000235:genetics; D001483:Base Sequence; D001692:Biological Transport; D002451:Cell Compartmentation; D003062:Codon; D004251:DNA Transposable Elements; D004269:DNA, Bacterial / Q000235:genetics; D004274:DNA, Recombinant; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D021382:Protein Sorting Signals / Q000235:genetics*; D001616:beta-Galactosidase / Q000235:genetics
D003062:Codon; D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D004274:DNA, Recombinant; D021382:Protein Sorting Signals; D000469:Alkaline Phosphatase; D001616:beta-Galactosidase