A highly efficient transposon mutagenesis system for the tomato pathogen Clavibacter michiganensis subsp. michiganensis.
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.
Kirchner
O
O
Fakultät für Biologie, Lehrstuhl für Mikrobiologie/Gentechnologie, Universität Bielefeld, Germany.
Gartemann
K H
KH
Zellermann
E M
EM
Eichenlaub
R
R
Burger
A
A
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
D004251:DNA Transposable Elements