Witte A, Reisinger G R, Säckl W, Wanner G, Lubitz W
Institute of Microbiology and Genetics, University of Vienna, Austria.
FEMS Microbiol Lett. 1998 Jul 1;164(1):159-67. doi: 10.1111/j.1574-6968.1998.tb13081.x.
Gene E-L, a chimeric lysis construct from bacteriophages phi X174 and MS2 lysis proteins E and L, respectively, was subjected to internal deletions to create a series of new E-L clones with altered lysis or killing properties. The lytic activities of the parental genes E. L. E-L and the internal truncated forms of E-L were investigated in this study to characterize the different lysis mechanisms, based on differences in the architecture of the different membrane spanning domains. Electron microscopy and release of marker enzymes for the cytoplasmic and periplasmic spaces revealed that two different lysis mechanisms can be distinguished depending on penetrating of the proteins either the inner membrane or the inner and outer membranes of Escherichia coli. Several candidates, which share efficient lysis properties, have biotechnological applications in terms of cell disruption.
基因E-L是一种嵌合裂解构建体,分别来自噬菌体phi X174和MS2的裂解蛋白E和L,对其进行内部缺失以创建一系列具有改变的裂解或杀伤特性的新E-L克隆。本研究研究了亲本基因E、L、E-L以及E-L的内部截短形式的裂解活性,以基于不同跨膜结构域的结构差异来表征不同的裂解机制。电子显微镜以及细胞质和周质空间标记酶的释放表明,根据蛋白质穿透大肠杆菌内膜还是内膜和外膜的情况,可以区分两种不同的裂解机制。几种具有高效裂解特性的候选物在细胞破碎方面具有生物技术应用。