Generation of readily transformable Bacillus licheniformis mutants

被引：71
作者：
Waschkau, Bianca [1 ]
Waldeck, Jens [1 ]
Wieland, Susanne [2 ]
Eichstaedt, Renee [2 ]
Meinhardt, Friedhelm [1 ]
机构：
[1] Univ Munster, Inst Mol Mikrobiol & Biotechnol, D-48149 Munster, Germany
[2] Henkel KGAA, VTB Enzymtechnol, D-40191 Dusseldorf, Germany
来源：
APPLIED MICROBIOLOGY AND BIOTECHNOLOGY | 2008年 / 78卷 / 01期
关键词：
Bacillus licheniformis; targeted gene deletion; restriction modification system; restriction-negative mutants; improved transformation; transformation;
D O I：
10.1007/s00253-007-1278-0
中图分类号：
Q81 [生物工程学（生物技术）]; Q93 [微生物学];
学科分类号：
071005 ; 0836 ; 090102 ; 100705 ;
摘要：
A set of mutants was generated by targeted deletion of the hsdR loci of two type I restriction modification systems ( RMS) identified in Bacillus licheniformis DSM13. Single as well as double knock-outs resulted in strains being readily transformable with plasmids isolated from Bacilli. Introduction of shuttle plasmids isolated from Escherichia coli was routinely possible when the double mutant B. licheniformis MW3 (Delta hsdR1, Delta hsdR2) was used in transformation experiments. Growth and secretion of extracellular enzymes were not affected in any of the mutants. Thus, along with an optimized transformation protocol, this study makes available an urgently needed transformation system for this industrially exploited species.
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页码：181 / 188
页数：8
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