Manoil C, Beckwith J
Proc Natl Acad Sci U S A. 1985 Dec;82(23):8129-33. doi: 10.1073/pnas.82.23.8129.
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.
我们构建了转座子Tn5的一个衍生物，它能产生由缺少信号肽的碱性磷酸酶（EC 3.1.3.1）与其他蛋白质的氨基末端序列融合而成的杂合蛋白。如果与碱性磷酸酶融合的蛋白质提供了促进输出的序列，从而弥补了缺失的碱性磷酸酶信号肽，这样的杂合蛋白就会具有碱性磷酸酶活性。与分泌型周质蛋白和复杂的细胞质膜蛋白的融合都导致了碱性磷酸酶活性。TnphoA融合有助于在蛋白质结构（如跨膜蛋白）中定位输出信号，以及鉴定分泌蛋白和跨膜蛋白的新染色体基因。
