油桐LACS4基因的克隆与原核及真核表达

发布时间：2018-01-30 01:07

本文关键词： 油桐长链酰基辅酶A合成酶原核表达真核表达　出处：《经济林研究》2017年02期 　论文类型：期刊论文

【摘要】：长链酰基辅酶A合成酶(LACS)在油脂代谢过程中发挥着重要的作用。以油桐近成熟种子为材料,在已构建的转录组数据库基础上,克隆得到一个油桐长链酰基辅酶A合成酶基因,在此基因c DNA全长序列的基础上,成功构建了油桐LACS4基因的真核表达和原核表达载体,其中,酵母表达载体p YES2-LACS4成功转入酿酒酵母菌株INVSc1中,并进行相关生长趋势分析;在大肠杆菌BL21(DE3)宿主细胞中,原核表达载体p ET30a-LACS4也成功诱导表达,并获得大约为74k Da的表观分子量的相应目的蛋白。
[Abstract]:Long-chain coenzyme A synthase (Lacs) plays an important role in lipid metabolism. The nearly mature seeds of Taulownia were used as materials and based on the constructed transcriptome database. A long chain acylcoenzyme A synthase gene was cloned and the eukaryotic expression and prokaryotic expression vector of LACS4 gene were successfully constructed on the basis of the full-length sequence of c DNA of the gene. Yeast expression vector p YES2-LACS4 was successfully transferred into Saccharomyces cerevisiae strain INVSc1 and the related growth trend was analyzed. The prokaryotic expression vector p ET30a-LACS4 was also successfully induced in the host cells of Escherichia coli BL21 DDE3. The corresponding target protein with an apparent molecular weight of about 74 kDa was obtained.
【作者单位】：中南林业科技大学经济林培育与保护省部共建教育部重点实验室;中南林业科技大学经济林育种与栽培国家林业局重点实验室;中南林业科技大学经济林培育与利用湖南省2011协同创新中心;
【基金】：湖南省重点研发计划“农业领域技术创新项目”(2016NK2147) 湖南省研究生科研创新项目(CX2016B327) 中南林业科技大学研究生科技创新基金(CX2016B15)
【分类号】：S794.3
【正文快照】： 油桐Vernicia fordii隶属大戟科Euphorbiaceae油桐属Vernicia Lour.,是我国四大木本油料树种之一[1-2]。油桐的种仁含油量高达60%,个别甚至超过70%,桐油的脂肪酸含量主要包含硬脂酸、软脂酸、桐酸、亚油酸、亚麻酸以及油酸[3],其中,桐酸是油桐所特有的脂肪酸,占总脂肪酸含量的

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