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FUNCTIONAL ANALYSIS OF OLEATE DESATURASE GENE EXPRESSION FROM BRASSICA JUNCEA

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Title FUNCTIONAL ANALYSIS OF OLEATE DESATURASE GENE EXPRESSION FROM BRASSICA JUNCEA
 
Creator SURESHA, G.S.
 
Contributor I. M. Santha
 
Subject acidity, genes, unsaturated fatty acids, fatty acids, planting, sowing, genetic structures, genomes, enzymes, biological phenomena
 
Description t-8043
Higher plants express one or more microsomal oleic acid desaturase (FAD2;
EC1.3.1.35) isoforms which catalyze the insertion of a double bond between carbons
12 and 13 of oleic acid at both the sn-1 and sn-2 positions of phosphatidylcholine. In
the present study, full length genomic and cDNA sequences of fad2 gene have been
isolated by using PCR approach from Brassica juncea. Analysis of genomic structure
of fad2 gene revealed that isolated genomic sequence (2526bp) contains 1238 bp 5’
untranslated region which is having a large single intron of 1081 bp, 143 bp 3’
untranslated region, and a CDS of 1155 bp. Analysis of cDNA sequence (1445 bp)
corresponding to genomic sequence (2526bp) revealed that isolated cDNA sequence
exactly corresponded with the genomic sequence containing 5’UTR of 147 bp, CDS of
1155 bp and 3’UTR of 143 bp. Southern blot analysis using partial fad 2 genomic
sequence as probe confirmed that there are atleast 2 copies of fad2 gene present in the
Brassica juncea genome. ClustalW alignment of deduced amino acid sequences of the
BjFAD2 protein together with the other plant fatty acid desaturases showed that all
these membrane-bound desaturases contained the three conserved histidine clusters
[HXXXH,HXXHH and HXXHH] that have been shown essential for desaturase
activity-acting as potential ligands for non-heme iron atoms. Phylogenetic analysis of
deduced amino acid sequences of BjFAD2 with other plant microsomal (FAD2) and
plastidial (FAD6) oleate desaturases showed that a BjFAD2 was positioned in a
subgroup with FAD2 enzymes that exhibit a house keeping pattern of expression.
Study of developmental expression of fad2 gene from developing seeds of Brassica
juncea through RT-PCR revealed that expression of fad2 gene was induced in early
stages of seed development (15 DAF), peaked in mid maturation stage (30 DAF) and
declined as seeds matured (45 DAF). Study of temperature dependent expression of
fad2 gene through RT-PCR revealed that expression of fad2 gene increased under
lower temperature treatments with incubation time as compared to the higher
 
Date 2017-01-03T13:41:51Z
2017-01-03T13:41:51Z
2008
 
Type Thesis
 
Identifier http://krishikosh.egranth.ac.in/handle/1/94143
 
Format application/pdf