title: A highly efficient transposon mutagenesis system for the tomato pathogen Clavibacter michiganensis subsp. michiganensis.
abstract: A transposon mutagenesis system for Clavibacter michiganensis subsp. michiganensis was developed based on antibiotic resistance transposons that were derived from the insertion element IS1409 from Arthrobacter sp. strain TM1 NCIB12013. As a prerequisite, the electroporation efficiency was optimized by using unmethylated DNA and treatment of the cells with glycine such that about 5 x 10(6) transformants per microg of DNA were generally obtained. Electroporation of C. michiganensis subsp. michiganensis with a suicide vector carrying transposon Tn1409C resulted in approximately 1 x 10(3) transposon mutants per pg of DNA and thus is suitable for saturation mutagenesis. Analysis of Tn1409C insertion sites suggests a random mode of transposition. Transposition of Tn1409C was also demonstrated for other subspecies of C. michiganensis.
authors.affiliation: Fakultät für Biologie, Lehrstuhl für Mikrobiologie/Gentechnologie, Universität Bielefeld, Germany.
authors.forename: O
authors.initials: O
authors.lastname: Kirchner
authors.forename: K H
authors.initials: KH
authors.lastname: Gartemann
authors.forename: E M
authors.initials: EM
authors.lastname: Zellermann
authors.forename: R
authors.initials: R
authors.lastname: Eichenlaub
authors.forename: A
authors.initials: A
authors.lastname: Burger
mesh_terms: D000192:Actinomycetales / Q000235:genetics* / Q000472:pathogenicity*; D004251:DNA Transposable Elements / Q000235:genetics; D018274:Electroporation / Q000379:methods; D005822:Genetic Vectors; D018551:Solanum lycopersicum / Q000382:microbiology*; D016254:Mutagenesis, Insertional / Q000379:methods*; D010935:Plant Diseases / Q000382:microbiology
chemical_list: D004251:DNA Transposable Elements