Molecular cloning of partial oleate desaturase gene from Brassica napus

The polyunsaturated fatty acids linoleate and α-linolenate are important membrane components and are the essential fatty acids of human nutrition. The major enzyme responsible for the synthesis of these compounds is the plant oleate desaturase of the endoplasmic reticulum, and its activity is controlled in Brassica napus by the fatty acid desaturation 2 (fad2) locus. We report the isolation of partial fad2 gene fragment from Brassica napus. A pair of primers were synthesized based on the conserved region of all plant fad 2 gene sequences available in NCBI database. Using these primers in PCR, 987 bp fragment was amplified from Brassica napus genomic DNA. After purification, the fragment was cloned into the pGEMT Easy vector and identified as a part of the gene encoding oleate desaturase. Comparison of the nucleotide sequence revealed similarity with other reported oleate desaturase gene sequences. It can be used to isolate full length gene/cDNA encoding oleate desaturase.