Cloning and Expression of Chitinase Gene Isolated from Insect Pathogenic Fungi, Beauveria bassiana in Escherichia coli
Khemika Songjang*[a], Tawee Donchai [a], Paranya Chaiyawat [a] and Christopher R. Meyer [b]* Author for corresponding; e-mail address: khemika@chiangmai.ac.th
Volume: Vol.33 No.3 (SEPTEMBER 2006)
Research Article
DOI:
Received: 15 Febuary 2006, Revised: -, Accepted: 21 July 2006, Published: -
Citation: Songjang K., Donchai T., Chaiyawat P. and Meyer C., Cloning and Expression of Chitinase Gene Isolated from Insect Pathogenic Fungi, Beauveria bassiana in Escherichia coli, Chiang Mai Journal of Science, 2006; 33(3): 347-355.
Abstract
A chitinase gene (1047 bp) was amplified from Beauveria bassiana BCC3653 genomic DNA by PCR technique using designed primers to generate 5’-EcoRI and 3’-HindIII ends. As this gene contains no intron, the PCR product was cloned into an expression vector, pSE420 resulting a recombinant plasmid, pDSM1. This recombinant plasmid was then transformed into Escherichia coli TOP10 cells. After confirming the DNA sequence of cloned PCR, gene expression was induced by IPTG. The chitinase activities were then investigated in gel supplemented with glycol chitin. Two chitinolytic activity bands were detected only in E. coli carrying pDSM1 suggesting the expression of cloned chitinase gene.