Development of An Asporogenic Mutant of Bacillus Licheniformis PWD-1.

[摘要] 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.

[发布机构] University:North Carolina State University

[效力级别] sigma F

[关键词] spoIIAC;sigma F;sporulation;bacillus licheniformis