title: A method to capture large DNA fragments from genomic DNA.
abstract: The gene capture technique is a powerful tool that allows the cloning of large DNA regions (up to 80 kb), such as entire genomic islands, without using restriction enzymes or DNA amplification. This technique takes advantage of the high recombinant capacity of the yeast. A "capture" vector containing both ends of the target DNA region must first be constructed. The target region is then captured by co-transformation and recombination in yeast between the "capture" vector and appropriate genomic DNA. The selected recombinant plasmid can be verified by sequencing and transferred in the bacteria for multiple applications. This chapter describes a protocol specifically adapted for Pseudomonas aeruginosa genomic DNA capture.
authors.affiliation: Laboratoire d'Ingénierie des Systèmes Macromoléculaires, UMR7255 - CNRS - Aix Marseille Université - IMM, 31 chemin Joseph Aiguier, 13402, Marseille cedex 20, France, ball@imm.cnrs.fr.
authors.forename: Geneviève
authors.initials: G
authors.lastname: Ball
authors.forename: Alain
authors.initials: A
authors.lastname: Filloux
authors.forename: Romé
authors.initials: R
authors.lastname: Voulhoux
mesh_terms: D000085:Acetates / Q000494:pharmacology; D004269:DNA, Bacterial / Q000235:genetics*; D005822:Genetic Vectors / Q000378:metabolism; D016680:Genome, Bacterial / Q000235:genetics*; D023281:Genomics / Q000379:methods*; D010957:Plasmids / Q000378:metabolism; D011550:Pseudomonas aeruginosa / Q000187:drug effects / Q000235:genetics*; D011995:Recombination, Genetic / Q000187:drug effects; D012441:Saccharomyces cerevisiae / Q000187:drug effects / Q000235:genetics; D014170:Transformation, Genetic / Q000187:drug effects
chemical_list: D000085:Acetates; D004269:DNA, Bacterial; C488804:lithium acetate