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. 1985 Dec;82(23):8129-33.
doi: 10.1073/pnas.82.23.8129.

TnphoA: a transposon probe for protein export signals

C Manoil, J Beckwith

TnphoA: a transposon probe for protein export signals

C Manoil et al. Proc Natl Acad Sci U S A. 1985 Dec.

Abstract

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.

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