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

spoIIAC;sigma F;sporulation;bacillus licheniformis

Greenhut, William Bradley ; Jason Shih, Committee Chair,Dr. Walter Dobrogosz, Committee Member,Hosni M. Hassan, Committee Member,Greenhut, William Bradley ; Jason Shih ; Committee Chair ; Dr. Walter Dobrogosz ; Committee Member ; Hosni M. Hassan ; Committee Member

University:North Carolina State University

关键词: spoIIAC;    sigma F;    sporulation;    bacillus licheniformis;   

Others  :  https://repository.lib.ncsu.edu/bitstream/handle/1840.16/1792/etd.pdf?sequence=1&isAllowed=y

美国|英语

来源: null

【 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.