Cloning and Expression of Acyl Carrier Protein Gene from Schizochytrium

Abstract:

Acyl carrier protein is an essential component involved in the biosynthesis of DHA (Docosahexaenoic Acid) via PKS (Polyketide synthase) pathway, which takes the growing acyl chain from one enzyme to another. One cDNA clone, with high homology of ACP, was isolated from Schizochytrium sp. FJU-512 cDNA library. The deduced amino acid sequence contained 142 residues with isoelectric point of 5.04 and had the 4′-phosphopantetheine prosthetic(4′-PP) binding site. The target fragment was digested with BamHⅠ/HindⅢand inserted into the expression vector pET-30a resulting in the plasmid pET-30a/acp. The recombinant vector was transformed into E.coli BL21(DE3) and induced by IPTG. SDS-PAGE analysis demonstrated that ACP was effectively expressed.