Development of An Asporogenic Mutant of Bacillus Licheniformis PWD-1.
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Date
2004-12-19
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Abstract
Bacillus licheniformis PWD-1 is a keratin-degrading, spore-forming bacterium isolated from a poultry waste digester. A mutant of B. licheniformis PWD-1, named B. licheniformis WBG-2, was developed that is deficient in sporulation. The mutation was created using the Splicing by Overlap-Extension PCR method (Gene SOE'ing) to create a 256bp deletion in the spoIIAC gene, which encodes an essential sporulation-specific sigma factor. In-vivo gene replacement was accomplished with the use of a temperature-sensitive plasmid that is able to integrate and excise from the B. licheniformis chromosome. PCR and DNA sequencing were used to confirm the deletion, while heat-treatment assays and electron microscopy verified the absence of spores. The mutant, while completely asporogenic, is able to expresses normal levels of keratinase as compared with B. licheniformis PWD-1.
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spoIIAC, sigma F, sporulation, bacillus licheniformis
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Degree
MS
Discipline
Microbiology
