Molecular Cloning and Sequence Analysis of Class II Chitinase Gene in Leymus chinensis

[Objective] The aim of this study was to clone Class Ⅱ chitinase gene in Leymus chinensis grown in saline land in Heilongjiang Province and analyze its sequence,which provided a foundation for further study on the biological function and application of chitinase gene. [Method] cDNA library of Leymus chinensis leaves were constructed,and its DNA sequence was determined or analyzed,while the homology of chitinase gene and amino acid sequence was compared with that in GenBank. [Result] One full length cDNA fragment with length of 996 bp was cloned from cDNA library of Leymus chinensis leaves. The length of ORF was 768 bp encoding 225 amino acids (GenBank accession number: EU344908). The encoding products lacked CBD and C-terminal extension region from the view of structure,but had structural characters of Class Ⅱ chitinase gene,which indicated that amino acid sequence had high homology compared with Class Ⅱ chitinase gene of rye and wheat. The constructed recombinant vector pQE-LcChi2 could express a protein of 27 kD through induction,which was consistent with the deduced encoding product of pQE-LcChi2 gene. [Conclusion] LcChi2 gene is an expression gene,which can express in E.coli.