A Highly Efficient Transposon Mutagenesis System for the Tomato Pathogen Clavibacter michiganensis subsp. michiganensis | Zendy

Oliver Kirchner | Zendy; KarlHeinz Gartemann | Zendy; Eva-Maria Zellermann | Zendy; Rudolf Eichenlaub | Zendy; A. Burger | Zendy

Open Access

A Highly Efficient Transposon Mutagenesis System for the Tomato Pathogen Clavibacter michiganensis subsp. michiganensis

Author(s) -

Oliver Kirchner,

KarlHeinz Gartemann,

Eva-Maria Zellermann,

Rudolf Eichenlaub,

A. Burger

Publication year - 2001

Publication title -

molecular plant-microbe interactions

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 1.565

H-Index - 153

eISSN - 1943-7706

pISSN - 0894-0282

DOI - 10.1094/mpmi.2001.14.11.1312

Subject(s) - transposon mutagenesis , transposable element , clavibacter michiganensis , biology , transposition (logic) , mutagenesis , dna , mutant , genetics , microbiology and biotechnology , pathogen , gene , linguistics , philosophy

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 × 10 6 transformants per μg of DNA were generally obtained. Electroporation of C. michiganensis subsp. michiganensis with a suicide vector carrying transposon Tn1409C resulted in approximately 1 × 10 3 transposon mutants per μg 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.

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