Isolation of delta-9 stearoyl-acp desaturase gene from Chlorella sp

Abstract

Delta-9-stearoyl-ACP desaturase (SACPD) gene is the key gene in converting the saturated stearoyl-ACP (C:18) to monounsaturated oleoyl-ACP (C 18:1) in fatty acid biosynthesis pathways. Manipulation of delta-9 stearoyl-ACP desaturase gene 1s an important step to enhance the accumulation of oleic acid level, so that, more monounsaturated oleic acid can be produced in Chiarella sp. The RT-PCR technique was used to isolate the SACPD cDNA clone from Chiarella sp. The total RNA was reverse transcribed with KPN-Tl 7 oligo-dT primer by using M-ML V reverse transcriptase. Four forward primers designed from the conserved regions of SACPD gene were used to amplify corresponding 3 '-end regions of the gene.