Design of Expression Cassettes for Inverted Repeat and Intron-Spliced Inverted Repeat of PATE Gene Silencing

Abstract

Palm oil of Elaeis guineensis Jacq. contains approximately 50% saturated and 50% unsaturated fatty acids. In the composition of saturated fatty acid, palmitic acid is the most abundant which is predominantly around 44% of the total fatty acids. The high level of accumulation of palmitic acid should be reduced in order to get a better dietary fatty acid and a more nutritional value oil in the market.The purpose in reducing palmitic acid content could be carried out by silencing palmitoyl-ACP thioesterase (PATE) gene activities. Primer pairs to amplify partial PATE fragments, intron and 3Untranslated Region (3UTR) were designed from PATE of Elaeis guineensis var tenera (GenBank DQ422858). PATE fragments, intron and 3UTR were constructed in two expression cassettes with inverted repeat (IR) and intron-spliced inverted repeat (ISIR) orientations to generate hair-pin RNAs (hpRNA). Combinations of restriction enzymes (RE) were used to confirm, the size, orientation and location of the inserts.