Schifferli D M, Beachey E H, Taylor R K
Department of Microbiology and Immunology, University of Tennessee, Memphis 38104.
Mol Microbiol. 1991 Jan;5(1):61-70. doi: 10.1111/j.1365-2958.1991.tb01826.x.
The 987P fimbrial gene cluster has been previously cloned as a 12 kb fragment from prototype strain 987. Gene products encoded by the whole clone were analysed by utilizing an in vivo system based on the induction of transcription by T7 RNA polymerase. The sensitivity of this technique permitted us to identify new proteins involved in 987P fimbriation. In total, eight proteins were detected, their genes (fasA to fasH) were mapped and their orientation of transcription determined. Several of the gene products demonstrated typical properties of exported proteins. Precursor and processed forms could be correlated after inhibiting protein transport with ethanol. The detection of enzymatically active fusion proteins after TnphoA (Tn5IS50L::phoA) mutagenesis supported and complemented these results. One protein encoded by the 12kb fragment was found not to be related to fimbriation but rather the product of the STla gene, identified as a component of a Tn1681-like transposon.
987P菌毛基因簇先前已从原型菌株987中作为一个12 kb的片段被克隆出来。利用基于T7 RNA聚合酶转录诱导的体内系统对整个克隆编码的基因产物进行了分析。该技术的敏感性使我们能够鉴定出参与987P菌毛形成的新蛋白质。总共检测到了8种蛋白质,对它们的基因(fasA至fasH)进行了定位,并确定了它们的转录方向。几种基因产物表现出输出蛋白的典型特性。在用乙醇抑制蛋白质转运后,可以将前体和加工形式联系起来。TnphoA(Tn5IS50L::phoA)诱变后对酶活性融合蛋白的检测支持并补充了这些结果。发现由12 kb片段编码的一种蛋白质与菌毛形成无关,而是STla基因的产物,该基因被鉴定为类Tn1681转座子的一个组成部分。