Intron-spliced gene silencing constructs for alteration of seed fatty acid composition by RNAI induced transgene silencing

With the impression to silence the endogenous fad 2–1 gene of Glycine max L. intron-spliced hairpin gene silencing (ihp) constructs was designed and produced using vicilin seed specific promoter. In this direction a 1273 bp cDNA sequence of Gmfad 2–1 was amplified using RT-PCR approach. The ihp construct, incorporating the 212 bp 3′ UTR of the fad 2–1 gene and were separated by iritron of the same gene in spliceable orientation. Efficacy of these constructs was tested in Arabidopsis thaliana. Confirmation of integration of transgene into host genome was confirmed by PCR using bar gene specific primer and further by southern hybridization.