Brassica napus is one of the most important oil crops in the world. The quality of rapeseed oil is determined by its fatty acids content. The saturated and unsaturated fatty acids biosynthesis pathway has many different stages which are controlled by various enzymes. Modification of the Brassica oil for the desired fatty acid profile through genetic manipulation necessitates identification and isolation of genes involved in the fatty acid biosynthesis. Erucic aicd (C22:1) is one of the most saturated fatty acids that affects rapeseed oil quality, when its amount increase to over 2 persen becomes undesiralae for human consumption. ? -ketoacyl-CoA synthase (KCS) was shown to be one of the key enzymes for biosynthesis of erucic acid. This enzyme, produeced by fae (fatty acid elongase), is involved in production of eicosenoic (C20: 1) and erucic acids (C22: 1) from C18 fatty acids. In this study we reported isolation, cloning and transformation of fae gene into B. napus. (For gene silencing and prevention of erucic acid synthesis antisense conctruct from fae gene was produced. Primer spicific design was done for gene isolation and its amplification. The fae gene (1521 bp) was isolated from genomic DNA by PCR technique. Digestion by restrictive enzymes (Hind III and EcoRI) was used to insure and ????? the fragment. Then, gene was cloned into pSK+ vector and sequenced with standard primers. Antisense construct from fae gene was cloned in cloning site of plant binary vector pBI121. This vector was then introduced to Agrobacterium tumefaciens to be used in plant transformation. Growth of transgenic plant on medium with kanamycin and its PCR shown that these plant were transformed. Dot blotting and Southern blot on transgenic plants were done. Silencing of fae gene by antisense construct be caused remarkable reduce in content of erucic acid in transformed plants of B. napus.
)