中国生物工程杂志

A chitosanase-producing bacterium S-1 was isolated from Lianyungang seaside. It was identified as Bacillus sp. based on its morphological, the physiological and biochemical properties and 16S rDNA sequence analysis. According to the information about chitosanases sequence deposited in NCBI, a pair of degenerated primers was designed and a partial sequence of chitosanase gene was obtained by polymerase chain reaction (PCR) using Bacillus sp. S-1 genome DNA as the template. A genome walking library was constructed followed as the protocol provided by CLONTECH Company. The flanking sequences of the 5' and 3' terminal was obtained by genome walking method and two-step PCR technique. After overlapped and confirmed, the full-length sequence of chitosanase from Bacillus sp. S-1 was achieved (Accession number: EU924147) and it contained 1362 bp coding 453 amino acids. The characterization of the deduced amino acid was analysis using the bioinformatics method.