Cloning and Characterization of Serine Protease Inhibitor Genes (SPIs) From Plants
| dc.contributor.advisor | Sumangala Bhat | |
| dc.contributor.author | D.A.Shirani | |
| dc.date.accessioned | 2016-10-17T13:33:46Z | |
| dc.date.available | 2016-10-17T13:33:46Z | |
| dc.date.issued | 2005 | |
| dc.description.abstract | Plant serine protease inhibitors (SPIs) can be effective in developing transgenic crop plants with resistance to insect attack. Such genes have few problems related to codon bias, RNA processing and protein stability. In the present study, an attempt was made to clone serine protease inhibitor genes from soybean (Glycine max), cowpea (Vigna unguiculata) and yard long bean (Vigna sesquipedalis). Using specific primers, genes encoding kunitz trypsin inhibitor of soybean (SKTI) and cowpea trypsin inhibitor (CpTI) from cowpea and yard long bean were amplified and cloned into pTZ57R/T vector. The clones were confirmed through PCR amplification and restriction digestion analysis. Further, the clones were sequenced and analyzed in silico. Cloned kunitz trypsin inhibitor of soybean showed 99 and 100 per cent homology with reported kunitz inhibitor genes of soybean at nucleotide and protein level respectively. The cloned CpTI of cowpea and yard long bean showed 99% homology with Phaseolus vulgaris trypsin proteinase inhibitor gene and published CpTI coding sequences (324bp). The deduced amino acid sequence of cloned cowpea CpTI showed 100% homology with Phaseolus vulgaris trypsin inhibitor whereas CpTI of yard long bean showed 98 percent homology. Further, to facilitate transformation of crop plants, soybean kunitz trypsin inhibitor and CpTI of cowpea and yard long bean were sub cloned into a plant transformation vector pHS100 at BamH1 and Xba1 restriction sites. The recombinant clones pHS-SBC1, pHS-CpC1 and pHS-YLBC3 were mobilized into Agrobacterium tumefaciens LBA4404 by triparental mating. The A. tumefaciens with recombinant clones pHS-SBC1 and pHS-CpC1 were used to transform tobacco and the presence of inserts in kanamycin resistant plants was checked through PCR with specific primers. The events have to be confirmed further and expression tested. | en_US |
| dc.identifier.uri | http://krishikosh.egranth.ac.in/handle/1/80703 | |
| dc.publisher | UAS, Dharwad | en_US |
| dc.research.problem | Cloning and Characterization of Serine Protease Inhibitor Genes (SPIs) From Plants | en_US |
| dc.sub | Plant Biochemistry | en_US |
| dc.subject | Plant Bio-Technology | en_US |
| dc.theme | Cloning and Characterization of Serine Protease Inhibitor Genes (SPIs) From Plants | en_US |
| dc.title | Cloning and Characterization of Serine Protease Inhibitor Genes (SPIs) From Plants | en_US |
| dc.type | Thesis | en_US |