Molecular cloning of a chitinase gene from Vibrio sp. KOPRI 11027

Abstract

The chiK gene encoding a chitinase ChiK was cloned from Vibrio sp. 98CJII027 using PCR method. The chiK open reading frame is 2553 bp long encoding a ChiK precursor protein, which contains 850 amino acids including a N-terminal signal peptide consisted of 21 amino acids. Using inverse PCR cloning method, we identified the 6.0 kb DNA fragment containing the chiK gene Subsequently, a 3042 bp of full chiK gene was cloned by PCR cloning, and then it was subcloned into pGEM T-easy vector, designated as pChiK. The recombinant E. coli π11109 clone harboring recombinant pChiK produced a clear halo around the colony in the colloidal chitin plates. The dedllced amino acid seqllences of ChiK revealed 89% identity with those of V&igrave

brio harveyi chitinase. Comparison of the conserved modllle reveals that chitinase of KOPRI 11027 strain(ChiK) belongs to the family 18 of gIycoside hydrolases.