title: Substrate and target sequence length influence RecTE(Psy) recombineering efficiency in Pseudomonas syringae.
abstract: We are developing a new recombineering system to assist experimental manipulation of the Pseudomonas syringae genome. P. syringae is a globally dispersed plant pathogen and an important model species used to study the molecular biology of bacteria-plant interactions. We previously identified orthologs of the lambda Red bet/exo and Rac recET genes in P. syringae and confirmed that they function in recombineering using ssDNA and dsDNA substrates. Here we investigate the properties of dsDNA substrates more closely to determine how they influence recombineering efficiency. We find that the length of flanking homologies and length of the sequences being inserted or deleted have a large effect on RecTE(Psy) mediated recombination efficiency. These results provide information about the design elements that should be considered when using recombineering.
authors.affiliation: Department of Plant Pathology and Plant-Microbe Biology, Cornell University, Ithaca, New York, United States of America.
authors.forename: Zhongmeng
authors.initials: Z
authors.lastname: Bao
authors.forename: Sam
authors.initials: S
authors.lastname: Cartinhour
authors.forename: Bryan
authors.initials: B
authors.lastname: Swingle
mesh_terms: D004277:DNA, Single-Stranded / Q000235:genetics / Q000378:metabolism; D005798:Genes, Bacterial / Q000235:genetics*; D005818:Genetic Engineering / Q000379:methods*; D044224:Pseudomonas syringae / Q000235:genetics*; D011995:Recombination, Genetic; D012689:Sequence Homology, Nucleic Acid
chemical_list: D004277:DNA, Single-Stranded