title: Rapid and efficient transposon mutagenesis of Bartonella henselae by transposome technology.
abstract: Molecular genetics are difficult to perform in Bartonella henselae, the causative agent of cat scratch disease and the vasculoproliferative disorders bacillary angiomatosis and bacillary peliosis. To elucidate the underlying bacterial pathogenic mechanisms, genetic manipulation of B. henselae is the method of choice. We describe how to perform transposon mutagenesis in B. henselae using transposome technology. B. henselae mutants revealed by this technique showed random transpositional insertion into the chromosome. In contrast to transposon mutagenesis by conjugational transfer, transposome technology allows transposon mutagenesis of early passaged Bartonella spp. with approximately 100-fold higher efficiency. The results show that transposome technique is a rapid, efficient and simple method to generate transposon mutants of B. henselae.
authors.affiliation: Institut für Medizinische Mikrobiologie und Krankenhaushygiene, Eberhard Karls Universität, D-72076 Tübingen, Germany.
authors.forename: Tanja
authors.initials: T
authors.lastname: Riess
authors.forename: Burt
authors.initials: B
authors.lastname: Anderson
authors.forename: Andrea
authors.initials: A
authors.lastname: Fackelmayer
authors.forename: Ingo B
authors.initials: IB
authors.lastname: Autenrieth
authors.forename: Volkhard A J
authors.initials: VA
authors.lastname: Kempf
mesh_terms: D018416:Bartonella henselae / Q000166:cytology / Q000235:genetics*; D015139:Blotting, Southern; D002460:Cell Line; D018920:Coculture Techniques; D004251:DNA Transposable Elements / Q000235:genetics*; D004269:DNA, Bacterial / Q000235:genetics / Q000378:metabolism; D015246:Deoxyribonuclease EcoRI / Q000378:metabolism; D015247:Deoxyribonuclease HindIII / Q000378:metabolism; D015252:Deoxyribonucleases, Type II Site-Specific / Q000378:metabolism; D006801:Humans; D016254:Mutagenesis, Insertional / Q000379:methods*; D009154:Mutation
chemical_list: D004251:DNA Transposable Elements; D004269:DNA, Bacterial; D015246:Deoxyribonuclease EcoRI; D015247:Deoxyribonuclease HindIII; C034510:CTCGAG-specific type II deoxyribonucleases; D015252:Deoxyribonucleases, Type II Site-Specific