Vol. 6 (2) : March-April 2015 issue
Green Farming Vol. 6 (2) : 341-345 ; March-April, 2015
Cloning of chitinase gene (Chit18-13) from Trichoderma reesei and its characterization
RAMESH METHRE1*, SUMANGALA BHAT2 and CHIDANAND A. RABINAL3
Department of Biotechnology, University of Agricultural Sciences, Dharwad - 580 005 (Karnataka)
Designation : 1,3Ph.D. Scholar *(rm.metre@gmail.com), 2Professor
Subject : Biotechnology, Molecular biology, Agriculture Microbiology and Cancer Res.
Paper No. : P-2371
Total Pages : 5
Received : 02 July 2014
Revised accepted : 13 August 2014
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Citation :
RAMESH METHRE, SUMANGALA BHAT and CHIDANAND A. RABINAL. 2015. Cloning of chitinase gene (Chit18-13) from Trichoderma reesei and its characterization. Green Farming Vol. 6 (2) : 341-345 ; March-April, 2015
ABSTRACT
The present study was conducted to clone endochitinase gene chit18-13, from Trichoderma reesei using gene specific primers. The amplicon of 1300bp size was amplified and cloned into pTZ57R/T vector. The sequence results showed 99 and 98 per cent homology with T. reesei chit18-13 endochitinase gene at nucleotide (accc.no.BK006086.1) and protein level (acc.no.DAA05861.1) in blastn and blastp programme. This protein has N-terminal signal peptide from 1-14 amino acids (MFFSKALAAAGLLA), catalytic domain from 31-259 amino acids and chitinase active sites from 163-171 amino acids (LDGVDLDNE). Phylogenetic analysis of cloned gene chit18-13 indicated that, this gene fall into different cluster and not related closely to other genes cloned in our laboratory from Trichoderm virens, Trichoderma viride and Trichoderma harzianum.
Key words :
Cloning, Endochitinase gene, Trichoderma reesei, Transformation.
