A highly efficient transposon mutagenesis system for the tomato pathogen Clavibacter michiganensis subsp michiganensis

被引：33

作者：

Kirchner, O ^{[1
]}

Gartemann, KH ^{[1
]}

Zellermann, EM ^{[1
]}

Eichenlaub, R ^{[1
]}

Burger, A ^{[1
]}

机构：

[1] Univ Bielefeld, Dept Microbiol Genetechnol, Fak Biol, D-33615 Bielefeld, Germany

来源：

MOLECULAR PLANT-MICROBE INTERACTIONS | 2001年 / 14卷 / 11期

关键词：

D O I：

10.1094/MPMI.2001.14.11.1312

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

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 jag 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 mug 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.

引用

页码：1312 / 1318

页数：7

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