S366 : Sequencing and cloning of two genes involved in the biosynthesis of Omega-3 in Rape
Thesis > Central Library of Shahrood University > Agricultural Engineering > MSc > 2017
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Abstarct: Plants can be used as sources for the production of omega-3 fatty acids, and this process is possible by genetic engineering of plant to have genes that encoding this biosynthetic pathway enzymes. Rapeseed can be a good candidate for research on the vegetable production of omega-3 fatty acids, given the high percentage of oil present in the seed, as well as the natural level of omega-3 fatty acids to omega-6 fatty acids in oil.
Despit that the common pathway for production of omega-3 is delta-6 desaturase pathway, this research with the goal of generating genetic data about enzymes that might be use for the production of omega-3 in canola was done. At first, Oil samples of rapeseed were extracted and using the method of gas chromatography, the type and amount of different fatty acids found in rapeseed oil were determined, then bioinformatics studies was done by comparing the sequences encoding the delta-6 desaturase and delta -6 elongase genes in other plant with the sequence of rapeseed genome. Subsequently, according to the results of bioinformatics studies, which resulted in no similar sequences encoding the delta-6 desaturase and delta-6 elongase genes in the rapeseed genome, the existence of genes encoding the delta-9 elongase pathway for the production of fatty acids omega-3 was examined in the rapeseed genome. Comparison of sequences encoding delta-9 elongase and delta-8 desaturase genes in databaxses with rapeseed genome resulted in detect of a series of sequences with similarity to delta-8 desaturase. consequently, with the comparison of different sequences encoding the delta-8 desaturase gene in plants, protected gene sequences of this gene were identified and used to design non-specific primers. RNA were extracted from leaf and seed tissues of rape and after cDNA synthesis, using the designed primers, Delta-8 desaturase sequence in the rapeseed genome were amplified. after electrophoresis and confirmation of expected length the amplified sequence were extracted from agarose gel and cloned in TA vector. Then, this transgenic vector was transferred to E. coli DH5a strain and after blue-white screening and successfully confirming the cloning sequencing was confirmed. At the end, the obtained sequence was compared with other delta-8 desaturase encoding sequences and thus was identified as a unique sequence.
Keywords:
#omega-3 #canola #delta-6 desaturase #delta-6 elongase #delta-9 elongase #delta-8 desaturase Link
Keeping place: Central Library of Shahrood University
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Keeping place: Central Library of Shahrood University
Visitor: