Recombineering and stable integration of the Pseudomonas syringae pv. syringae 61 hrp/hrc cluster into the genome of the soil bacterium Pseudomonas fluorescens Pf0-1.
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.
Thomas
William J
WJ
Department of Botany and Plant Pathology, Oregon State University, Corvallis, OR 97331, USA.
Thireault
Caitlin A
CA
Kimbrel
Jeffrey A
JA
Chang
Jeff H
JH
D003001:Cloning, Molecular; D005818:Genetic Engineering / Q000379:methods*; D016680:Genome, Bacterial; D011551:Pseudomonas fluorescens / Q000235:genetics*; D044224:Pseudomonas syringae / Q000235:genetics*; D011995:Recombination, Genetic; D014026:Nicotiana / Q000382:microbiology