African Journal of
Agricultural Research

  • Abbreviation: Afr. J. Agric. Res.
  • Language: English
  • ISSN: 1991-637X
  • DOI: 10.5897/AJAR
  • Start Year: 2006
  • Published Articles: 6574

Full Length Research Paper

Isolation and cloning of ech36 gene from Trichoderma harzanium

Radheshyam Sharma* and Sumangala Bhat
Institute of Agri-Biotechnology (IABT), College of Agriculture, University of Agricultural Sciences, AC, Dharwad-580 005 Karnataka, India. 
Email: [email protected]

  •  Accepted: 24 May 2013
  •  Published: 06 June 2013

Abstract

A lab experiment was conducted to screen for the presence of ech36 gene in 80 isolates of Trichoderma.Further, using gene specific primers ech36 gene were cloned into pTZ57R/T from Trichoderma harzaniumIABT1042. The clone was confirmed through PCR amplification and restriction analysis. The clone were sequenced and analyzed for homology at nucleotide and protein level to find out conserved domains of the putative gene and protein. The gene encoding endochitinase from the species have 99 and 100% homology with reported sequences both at nucleotide and protein level. The cloned ech36 has a size of 1212 bp, of which 12 bp corresponds to the 3' untranslated region, with a 1030 bp open reading frame. The amino acid sequence of gene has signal peptide sequence ranges from1 to 25. The nucleotide sequence analysis using GENETOOL software revealed presence of three exon and introns and has unique restriction sites for HindIII,BglII, and HaeIII at 908, 387 and 672 positions respectively

 

Key words: Trichoderma harzanium, ech36, protein.