中华绒螯蟹Δ9-脂肪酸去饱和酶基因克隆和真核表达系统的构建
发布时间：2019-08-05 08:22
【摘要】：Δ9-脂肪酸去饱和酶是多不饱和脂肪酸合成途径的关键酶之一,可催化脂肪酸链上特定位置形成双键。本研究在已克隆到中华绒螯蟹(Eriocheir sinensis)的Δ9-脂肪酸去饱和酶(Δ9 fatty acyl-Co A desaturase,Δ9-FAD)基因基础上,为进一步了解其功能,根据中华绒螯蟹Δ9-FAD基因c DNA序列(accession number:JQ693685)以及真核表达载体p PIC3.5K,设计引物并在其上下游分别加入Bam HⅠ和Eco RⅠ酶切位点;利用反转录PCR(RT-PCR)技术,克隆其开放阅读框(open reading frame,ORF)片段,构建重组质粒p PIC3.5k-Δ9-FAD;利用电穿孔仪将经限制性内切酶SacⅠ线性化后的重组质粒p PIC3.5k-Δ9-FAD转入毕赤酵母(Pichia pastoris)GS115菌株中。经筛选与PCR验证,得到含有重组质粒p PIC3.5k-Δ9-FAD的毕赤酵母转化株。本实验成功构建了中华绒螯蟹Δ9-脂肪酸去饱和酶的毕赤酵母表达系统,为深入研究Δ9-脂肪酸去饱和酶的功能做了基础性的工作。
[Abstract]:螖 9-fatty acid desaturase is one of the key enzymes in the synthesis of polysaturated fatty acids, which can catalyze the formation of double bonds at specific positions in fatty acid chains. In this study, the 螖 9-fatty acid desaturase (螖 9 fatty acyl-Co A desaturase, 螖 9-FAD) gene of (Eriocheir sinensis) was cloned into Eriocheir sinensis (Eriocheir sinensis). In order to further understand its function, primers were designed according to the c DNA sequence (accession number:JQ693685) of the 螖 9-FAD gene and the eukaryotic expression vector p PIC3.5K, and Bam H 鈪，
本文编号：2523025
[Abstract]:螖 9-fatty acid desaturase is one of the key enzymes in the synthesis of polysaturated fatty acids, which can catalyze the formation of double bonds at specific positions in fatty acid chains. In this study, the 螖 9-fatty acid desaturase (螖 9 fatty acyl-Co A desaturase, 螖 9-FAD) gene of (Eriocheir sinensis) was cloned into Eriocheir sinensis (Eriocheir sinensis). In order to further understand its function, primers were designed according to the c DNA sequence (accession number:JQ693685) of the 螖 9-FAD gene and the eukaryotic expression vector p PIC3.5K, and Bam H 鈪，
本文编号：2523025
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