CLONING AND FUNCTIONAL CHARACTERIZATION OF ENDOCHITINASE GENE FROM TRICHODERMA SPP.
| dc.contributor.advisor | SUMANGALA BHAT | |
| dc.contributor.author | GANAPATI BHAT | |
| dc.date.accessioned | 2019-06-20T11:28:50Z | |
| dc.date.available | 2019-06-20T11:28:50Z | |
| dc.date.issued | 2007-08-30 | |
| dc.description.abstract | The present study was conducted to isolate Thchoderma spp. from different soil samples, to screen them for chitinase activity using glycol chitin plate assay, to clone full-length genes encoding endochitinase from Trichoderma species and expression of cloned endochitinase gene in Saccharomyces cerevisiae. During the study 44 Trichoderma isolates were obtained from 48 soil sample collected. Among the thirty three isolates tested for chitinase activity (glycol chitin plate assay), isolates T. virens lABT 1010, T. konlngiil^Ell^l^ and T. poiysporumlk^l were efficient producers of chitinase enzyme. Further, using specific primers, genes encoding endochitinase chiMS (1.6kb) from T viride and T. harzianum (1.5kb) were cloned into pTZ57R/T vector. The clones were confirmed through PGR amplification and restriction analysis. The clones were sequenced and analyzed for homology at nucleotide and protein level to find out conserved domain of protein. Genes encoding endochitinase from both the species have 95 % and 99.5% homology with reported sequence both at nucleotide and protein level. Phylogenetic analysis of cloned genes indicated that both the genes fall in same cluster and closely related to T harzianum endochitinase gene. Endochitinase gene cloned from T. v/re/75 (cloned earlier) was expressed in S. cerevisiae using pYES2/cr vector. Substrate conversion assay using cell lysate indicated 9.8 times more activity in recombinant clones compared to control. Further, cell lysates of recombinant clones significantly inhibited the growth of Scierotium roifsiL In addition, reduced expression of the same gene was observed when 173bp upstream sequence was removed. | en_US |
| dc.identifier.other | TH8743 | |
| dc.identifier.uri | http://krishikosh.egranth.ac.in/handle/1/5810109378 | |
| dc.language.iso | en | en_US |
| dc.pages | 151 | en_US |
| dc.publisher | University of Agricultural Sciences GKVK, Banglore | en_US |
| dc.sub | Biotechnology | en_US |
| dc.subject | null | en_US |
| dc.theme | CLONING AND FUNCTIONAL CHARACTERIZATION OF ENDOCHITINASE GENE FROM TRICHODERMA SPP. | en_US |
| dc.these.type | M.Sc | en_US |
| dc.title | CLONING AND FUNCTIONAL CHARACTERIZATION OF ENDOCHITINASE GENE FROM TRICHODERMA SPP. | en_US |
| dc.type | Thesis | en_US |