Molecular cloning of partial genomic and cDNA sequence of oleate desaturase gene from Brassica juncea.

摘要

In plants, the endoplasmic reticulum (ER)-associated oleate desatuarse (FAD 2) is the key enzyme responsible for the production of linoleic acid in non-photosynthetic tissues. In the present study, we isolated a partial genomic and cDNA sequences encoding microsomal oleate desaturase (FAD2) using a PCR approach. A pair of primers were synthesized based on the known plant fad2 gene sequences available in NCBI database. Using these primers in genomic PCR and RT PCR, 987 bp fragments were amplified from Brassica juncea. After purification, the fragments were cloned into the pGEMT Easy vector and sequenced. Isolated both genomic and cDNA sequences were identified as a part of the gene encoding oleate desaturase. Both genomic and cDNA fragments were 100% identical and showed that isolated genomic fragment does not contain any intron. Comparison of the nucleotide sequence revealed similarity with other reported oleate desaturase gene sequences. Isolated fad2 fragments could be a novel target for genetic manipulation of fad2 gene for desired oil quality in Brassica.