Molecular characterization and expression of SiFad1 in the sea urchin (Strongylocentrotus intermedius).

Fatty acid desaturases (Fads) are a key enzyme in the process of biosynthesis of highly unsaturated fatty acids (HUFAs). In this study, we cloned the full-length sequence of the SiFad1 gene (SiFad1) and analyzed its expression profiles during different developmental stages and in different tissues of Strongylocentrotus intermedius. The full-length cDNA of SiFad1 is composed of 1086 bp, with a putative open reading frame of 885 bp encoding a polypeptide of 294 amino acid (AA) residues. The predicted molecular mass of SiFad1 is 34.67 kDa and its theoretical pI is 8.41. The presence of conserved motifs including three histidine boxes (HXXXH, HXXHH, XXXHH), a FA_desaturases domain and three transmembrane domains suggests that SiFad1 belongs to the microsomal fatty acid desaturases family. Its tissue distribution showed that the highest expression of SiFad1 is in the intestine and the weakest expression is in Aristotle's lantern of S. intermedius. Time-course expression measurements in different developmental stages showed the highest expression of SiFad1 occurs in the gastrula and the weakest expression in the juvenile sea urchin. Knock-down of SiFad1 by specific siRNA revealed that the significantly depressed expression of Elovl5 had decreased in the coelomocytes, intestines and gonads at 24 h post transfection, indicating that the downstream target gene of SiFad1 is Elovl5 and SiFad1 and Elovl5 have positive regulatory effects. When we examined the changes in fatty acids in the gonads before and after interference, the results showed that after 24 h of interference, the content of C20:4n-6 produced by SiFad1 had decreased. Taken together, these results will enable us to understand the role of SiFad1 in fatty acid anabolism, which will help us to understand the fatty acid synthesis pathways and regulatory mechanisms of Strongylocentrotus intermedius and provide a theoretical experimental basis for improving the ability of sea urchins to synthesize fatty acids and cultivating sea urchins of higher quality and nutritional value.

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