摘要
The marine microalga Nannochloropsis sp.contains various elongases and desaturases that are critical for biosynthesis of polyunsaturated fatty acids.A full-length cDNA encoding a long-chain fatty acid elongase,named Ns FAE,was cloned from Nannochloropsis sp..The open reading frame of Ns FAE(GenBank accession no.MF680548)consisted of 1068 bp and encoded a predicted protein of 355 amino acids with molecular mass 38.8 k Da.The deduced polypeptide showed 43%–44%identity to fatty acyl elongases from other algae.RT-PCR experiments indicated that the Ns FAE gene exhibited the highest expression in Nannochloropsis sp.at 72 h(i.e.,during the third growth stage)and the expression was significantly lower in the other four growth stages.Plasmid p Ns FAE-CRISPR and a recombinant DNA fragment(ADH1p-Ns FAE-CYCt)were transformed into Saccharomyces cerevisiae strain BY4742 using the CRISPR-Cas system.Yeast transformants containing Ns FAE produced three fatty acids not normally present in wild-type BY4742-linoleic acid,linolenic acid and eicosadienoic acid-indicating that Ns FAE encodes a functional elongase enzyme.
The marine microalga Nannochloropsis sp. contains various elongases and desaturases that are critical for biosynthesis of polyunsaturated fatty acids. A full-length cDNA encoding a long-chain fatty acid elongase, named Ns FAE, was cloned from Nannochloropsis sp.. The open reading frame of Ns FAE(GenBank accession no. MF680548) consisted of 1068 bp and encoded a predicted protein of 355 amino acids with molecular mass 38.8 kDa. The deduced polypeptide showed 43%–44% identity to fatty acyl elongases from other algae. RT-PCR experiments indicated that the Ns FAE gene exhibited the highest expression in Nannochloropsis sp. at 72 h(i.e., during the third growth stage) and the expression was significantly lower in the other four growth stages. Plasmid p Ns FAE-CRISPR and a recombinant DNA fragment(ADH1 p-Ns FAE-CYCt) were transformed into Saccharomyces cerevisiae strain BY4742 using the CRISPR-Cas system. Yeast transformants containing Ns FAE produced three fatty acids not normally present in wild-type BY4742-linoleic acid, linolenic acid and eicosadienoic acid-indicating that Ns FAE encodes a functional elongase enzyme.
基金
supported by the Basic Scientific Fund for National Public Research Institutes of China (No. 2016Q07)
