Please use this identifier to cite or link to this item: http://krishikosh.egranth.ac.in/handle/1/5810039145
Authors: RICHARD LALROPUI
Advisor: Dr. H. M. VAMADEVAIAH
Title: DEVELOPMENT OF PTGS CONSTRUCT FOR THE SILENCING OF 9 DESATURASE GENE IN COTTON
Publisher: University of Agricultural Sciences Dharwad
Language: en
Type: Thesis
Pages: 99
Agrotags: null
Keywords: DEVELOPMENT OF PTGS CONSTRUCT FOR THE SILENCING OF ∆9 DESATURASE GENE IN COTTON SEED OIL
Abstract: Cotton (Gossypium spp.) is primarily grown for fibre production and it is also the world’s fifth largest source of vegetable oil. Cottonseed is a source of relatively high-quality protein (23%) with the addition of 21% oil. However, efficiency of the nutrient-rich resource for food and industrial purposes is being hampered by the presence of high palmitic and linoleic acid that form saturated fatty acid and later is thermostably polyunsaturated fatty acid. PTGS/RNAi is a novel gene regulatory mechanism that limits the transcript level by either suppressing transcription or by activating a sequence-specific RNA degradation process. Post transcriptional silencing of ∆9 desaturase gene was envisaged as a way for blocking the fatty acid biochemical pathway to increase the stearic acid production in the transformants. PTGS is a specific RNA degradation mechanism of any organism that takes care of aberrant unwanted excess of its own or foreign RNA intracellularly, in a homology dependent manner. In the present study, an effort was made to develop construct for the silencing of ∆9 desaturase gene, and analyse its expression in tobacco plants. Cotton ovules after 30 days pollination were collected and 600 bp DNA fragment was amplified using ∆9 desaturase gene specific primer, cloned into pTZ57R/T and confirmed by sequencing. The analysis of the sequence revealed maximum homology of 100 percent at nucleotide in the database (AJ132636.1). Partial ∆9 desaturase gene was cloned in sense, antisense and both (sense/antisense) orientation in generic ihp vector to develop PTGS constructs. The expression cassette carrying insert from generic ihp vector was then subcloned into plant transformation vector pCAMBIA 1305.1 to facilitate plant transformation. The recombinant clones were then mobilized into Agrobacterium tumefaciens LBA4404 by tri parental mating with E.coli pPRK2013 as helper strain and then transformed into tobacco plants. Callus was analyzed through histochemical analysis of gus gene expression against control plants.
Subject: Biotechnology
Theme: DEVELOPMENT OF PTGS CONSTRUCT FOR THE SILENCING OF ∆9 DESATURASE GENE IN COTTON SEED OIL
These Type: M.Sc
Issue Date: 2014-07
Appears in Collections:Thesis

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