油茶脂肪酸β氧化多功能蛋白基因的克隆与原核表达

发布时间：2018-03-16 13:27

本文选题：油茶　切入点：多功能蛋白　出处：《分子植物育种》2016年06期 　论文类型：期刊论文

【摘要】：油茶是中国重要的木本食用油料树种,在油茶种子的发育过程中,涉及到一系列调控代谢过程的酶和基因,脂肪酸β氧化多功能蛋白(MFP)是β氧化系统上的一类酶,它完成催化β-氧化途径中的水合与脱氢过程。本研究以国审油茶品种‘华硕’种子为材料,在已构建的转录组和表达谱数据库的基础之上,采用RACE技术,克隆到油茶脂肪酸β氧化多功能蛋白基因的cDNA全长序列,命名为CoMFP(Gen Bank登录号为KJ910342)。序列分析表明,该基因cDNA全长为2 541 bp,含有2 178 bp的开放读码框,编码725个氨基酸,分子量为79.033 3 kD,理论等电点pI为9.11,具有六个比较明显的跨膜区,不仅存有反式烯酰CoA水合酶(ECH)信号序列、3羟酰Co A脱氢酶(HCDH)信号序列、ECH基序和磷酸结合基序,也具有相当保守的盐桥、活性位点及五个亚结构域。成功地构建了原核表达载体并在宿主细胞BL21(DE3)上成功诱导表达,获得表观分子量约为79 kD的相应目的蛋白。
[Abstract]:Camellia oleifera is an important woody edible oil species in China. During the development of Camellia oleifera seeds, it involves a series of enzymes and genes that regulate the metabolic process. It completed the hydration and dehydrogenation process in the catalytic 尾 -oxidation pathway. In this study, the seeds of Asus, a Chinese Camellia oleifera cultivar, were used as materials, on the basis of the constructed transcriptome and expression profile database, RACE technique was used. The full-length cDNA sequence of the fatty acid 尾 oxidation multifunctional protein gene of Camellia oleifera was cloned and named as CoMFP(Gen Bank accession number KJ910342. The sequence analysis showed that the cDNA length of the gene was 2 541 BP, containing an open reading frame of 2 178 BP and encoding 725 amino acids. The molecular weight is 79.033 3 KD, and the theoretical isoelectric point Pi is 9.11. There are six obvious transmembrane regions. Not only are there three hydroxyl CoA dehydrogenase (HCDH) signal sequences, but also the phosphoric acid binding motifs. The prokaryotic expression vector was successfully constructed and induced on the host cell BL21DDE3. The corresponding target protein with an apparent molecular weight of about 79 KD was obtained.
【作者单位】：中南林业科技大学经济林培育与保护教育部重点实验室经济林育种与栽培国家林业局重点实验室;国家林业局桉树研究开发中心;广西林业科学研究院;华中师范大学生命科学学院;
【基金】：国家自然科学基金(31070603) 湖南省自然科学基金(14JJ2104) 中南林业科技大学青年基金重点项目(QJ2011008A)共同资助
【分类号】：S794.4

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