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A New Simple Method to Introduce Marker-free Deletions in the Bacillus Subtilis Genome

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Date 2010 Jan 9
PMID 20057169
Citations 25
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Abstract

A genetic tool to introduce marker-free deletions is essential for multiple manipulations of genomes. We report a simple and efficient method to create marker-free deletion mutants of Bacillus subtilis through transformation with recombinant PCR products, using the Escherichia coli mazF gene encoding an endoribonuclease that cleaves free mRNAs as a counter-selection tool. Our method will be applicable to any bacterium in which introduction of the mazF cassette into the genome by double crossover homologous recombination is possible.

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