Hinton J C, Salmond G P
Department of Biological Sciences, University of Warwick, Coventry, UK.
Mol Microbiol. 1987 Nov;1(3):381-6. doi: 10.1111/j.1365-2958.1987.tb01946.x.
Soft rot Erwinia species secrete a range of enzymes into the extracellular environment. Therefore, the genetically amenable Erwinia system is a useful model for the study of protein secretion by Gram-negative bacteria. We have used a lambda-sensitive derivative of Erwinia carotovora subspecies carotovora (Ecc) and the transposon TnphoA, to isolate a range of extracellular enzyme mutants. The use of TnphoA provides an enrichment for extracellular enzyme mutants over other transposon-based systems. In these mutants, the alkaline phosphatase activity of the hybrid protein is found in the periplasm, and is under the control of the Ecc promoters. Three TnphoA-induced extracellular enzyme mutants were studied in detail. One proved to be an enzyme structural gene mutant, whilst the other two appeared to be secretory mutants.
软腐欧文氏菌属细菌会向细胞外环境分泌一系列酶。因此，遗传上易于操作的欧文氏菌系统是研究革兰氏阴性菌蛋白质分泌的有用模型。我们利用胡萝卜软腐欧文氏菌胡萝卜软腐亚种（Ecc）的一种对λ敏感的衍生物和转座子TnphoA，分离出了一系列细胞外酶突变体。与其他基于转座子的系统相比，使用TnphoA能富集细胞外酶突变体。在这些突变体中，杂合蛋白的碱性磷酸酶活性存在于周质中，并受Ecc启动子的控制。对三个TnphoA诱导的细胞外酶突变体进行了详细研究。其中一个被证明是酶结构基因突变体，而另外两个似乎是分泌基因突变体。
