CLONING, SEQUENCING AND EXPRESSION OF KERATINASE GENE FROM INDIGENOUS Bacillus licheniformis N5 STRAIN
Abstract
DNA fragment coding for keratinase (Ker) gene from local isolate, Bacillus licheniformis N5 was generated using designed primers and polymerase chain reaction (PCR) technology. It was cloned into pJET1.2 vector and expressed into E. coli DH5α. Transformants acquiring desired gene was confirmed either by PCR or by screening the gene expression on selective medium. E. coli transformant harboring recombinant plasmid named Ker-NRC-1 was selected and the nucleotide sequence of ker gene was deposited in GenBank (GenBank accession number: KJ642621.1).The complete CDS of keratinase was consists of 1140 bp representing 379 amino acids equivalent to 38.8 kDa. Up to 55% increase of keratinase activity was observed among E. coli recombinant strains comparing with Bacillus licheniformis N5 donor strain. Conclusions: Bacillus licheniformis keratinase was cloned and successfully expressed using T7 promoter in E coli. The recombinant plasmid could be used to insert the desired gene into other bacterial hosts including Bacillus strains to obtain a superior keratinase producer strain which could be potentially used in biotechnological application
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