【摘要】：Δ9-脂肪酸去飽和酶是多不飽和脂肪酸合成途徑的關(guān)鍵酶之一,可催化脂肪酸鏈上特定位置形成雙鍵。本研究在已克隆到中華絨螯蟹(Eriocheir sinensis)的Δ9-脂肪酸去飽和酶(Δ9 fatty acyl-Co A desaturase,Δ9-FAD)基因基礎(chǔ)上,為進(jìn)一步了解其功能,根據(jù)中華絨螯蟹Δ9-FAD基因c DNA序列(accession number:JQ693685)以及真核表達(dá)載體p PIC3.5K,設(shè)計引物并在其上下游分別加入Bam HⅠ和Eco RⅠ酶切位點;利用反轉(zhuǎn)錄PCR(RT-PCR)技術(shù),克隆其開放閱讀框(open reading frame,ORF)片段,構(gòu)建重組質(zhì)粒p PIC3.5k-Δ9-FAD;利用電穿孔儀將經(jīng)限制性內(nèi)切酶SacⅠ線性化后的重組質(zhì)粒p PIC3.5k-Δ9-FAD轉(zhuǎn)入畢赤酵母(Pichia pastoris)GS115菌株中。經(jīng)篩選與PCR驗證,得到含有重組質(zhì)粒p PIC3.5k-Δ9-FAD的畢赤酵母轉(zhuǎn)化株。本實驗成功構(gòu)建了中華絨螯蟹Δ9-脂肪酸去飽和酶的畢赤酵母表達(dá)系統(tǒng),為深入研究Δ9-脂肪酸去飽和酶的功能做了基礎(chǔ)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，

本文編號：2523025