Department of Botany and Plant Pathology, Oregon State University, Corvallis, OR 97331, USA.
Plant J. 2009 Dec;60(5):919-28. doi: 10.1111/j.1365-313X.2009.03998.x. Epub 2009 Aug 13.
Many Gram-negative bacteria use a type III secretion system (T3SS) to establish associations with their hosts. The T3SS is a conduit for direct injection of type-III effector proteins into host cells, where they manipulate the host for the benefit of the infecting bacterium. For plant-associated pathogens, the variations in number and amino acid sequences of type-III effectors, as well as their functional redundancy, make studying type-III effectors challenging. To mitigate this challenge, we developed a stable delivery system for individual or defined sets of type-III effectors into plant cells. We used recombineering and Tn5-mediated transposition to clone and stably integrate, respectively, the complete hrp/hrc region from Pseudomonas syringae pv. syringae 61 into the genome of the soil bacterium Pseudomonas fluorescens Pf0-1. We describe our development of Effector-to-Host Analyzer (EtHAn), and demonstrate its utility for studying effectors for their in planta functions.
许多革兰氏阴性菌使用 III 型分泌系统（T3SS）与宿主建立联系。T3SS 是将 III 型效应蛋白直接注射到宿主细胞中的导管，这些蛋白在宿主细胞中操纵宿主以利于感染细菌。对于与植物相关的病原体，III 型效应蛋白的数量和氨基酸序列的变化以及它们的功能冗余性使得研究 III 型效应蛋白具有挑战性。为了减轻这一挑战，我们开发了一种将单个或特定组合的 III 型效应蛋白稳定递送到植物细胞中的方法。我们使用重组和 Tn5 介导的转座分别克隆和稳定整合丁香假单胞菌 pv. syringae 61 的完整 hrp/hrc 区到土壤细菌荧光假单胞菌 Pf0-1 的基因组中。我们描述了我们开发的效应物到宿主分析器（EtHAn），并展示了其用于研究效应物在植物体内功能的效用。
