Laboratory of Clinical Research on Infectious Diseases, International Research Center for Infectious Diseases, Research Institute for Microbial Diseases, Osaka University, Osaka, Japan.
Biochem Biophys Res Commun. 2012 Oct 19;427(2):242-7. doi: 10.1016/j.bbrc.2012.09.018. Epub 2012 Sep 17.
The type III secretion system (T3SS) of gram-negative bacteria involves dedicated protein translocation machinery that directly injects proteins into target cells. Pathogenic bacteria already benefit from this unique system. The successful functional cloning of this useful tool into non-pathogenic bacteria would help establish novel clinical and basic biotechnology strategies in areas such as vaccine administration, the development of screening systems for anti-T3SS drugs and the target-specific delivery of bioactive compounds. In this study, we successfully cloned the Vibrio parahaemolyticus T3SS1 genetic locus into a non-pathogenic Escherichia coli K-12 strain. Assays performed here revealed that the T3SS1 cloned into the E. coli K-12 strain has the ability to translocate V. parahaemolyticus T3SS1 secreted proteins. Importantly, we also observed this system to allow the E. coli K-12 strain to inject foreign protein, as well as the V. parahaemolyticus T3SS effector, into cultured cells. These results demonstrate a prospective useful tool with experimental and therapeutic applications.
革兰氏阴性菌的 III 型分泌系统（T3SS）涉及专门的蛋白质转运机制，可直接将蛋白质注入靶细胞。致病性细菌已经受益于这种独特的系统。成功地将这个有用的工具的功能性克隆到非致病性细菌中，将有助于在疫苗接种、抗 T3SS 药物筛选系统的开发以及生物活性化合物的靶向特异性递送上建立新的临床和基础生物技术策略。在这项研究中，我们成功地将副溶血弧菌 T3SS1 基因座克隆到非致病性大肠杆菌 K-12 菌株中。这里进行的测定表明，克隆到大肠杆菌 K-12 菌株中的 T3SS1 具有转运副溶血弧菌 T3SS1 分泌蛋白的能力。重要的是，我们还观察到该系统允许大肠杆菌 K-12 菌株将外源蛋白以及副溶血弧菌 T3SS 效应物注入培养细胞。这些结果表明该系统具有实验和治疗应用的有前景的有用工具。
