Title: Cloning of gene celC of Bacillus sp.CY1-3 strain and its expression in Escherichia coli
Abstract:A genomic library of Bacillus sp.CY1-3 strain was constructed in Escherichia coli strain DH5α by ligating BamHⅠ-digested pBluSKM vector with 2-7kb Sau3AⅠ-digested genomic DNA from Bacillus sp.CY1-3 s...A genomic library of Bacillus sp.CY1-3 strain was constructed in Escherichia coli strain DH5α by ligating BamHⅠ-digested pBluSKM vector with 2-7kb Sau3AⅠ-digested genomic DNA from Bacillus sp.CY1-3 strain.Six clones expressing carboxymethylcellulase(CMCase) activity were isolated from nearly 100000 clones by screening the library with Congo red dying.These clones were identified to be the same clone by restriction endonuclease digestion,and it was designated as pGJc-1.The 2-7kb fragment contained a 1593bp ORF,named celC,which codes 531 amino acid residues.Compared the amino acid sequence of CelC with those available in GenBank,it was highly identical to endo-β-1,4-glucanase from Bacillus strains.SDS-PAGE analysis showed that protein expressed from gene celC in E.coli had cellulase activity,and its molecular weight was 58.0ku approximately.Read More
Publication Year: 2006
Publication Date: 2006-01-01
Language: en
Type: article
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