论文部分内容阅读
长链酰基辅酶A合成酶(long chain acyl-Co A synthetase,LACS)是油脂代谢的重要催化酶。本研究采用RT-PCR技术,从花生(Arachis hypogaea L.)克隆到LACS1(Gen Bank登录号:KT932703),分析了该基因的结构组成,预测编码氨基酸与其他植物的同源性,采用实时Real-Time PCR技术对LACS1的组织表达进行研究。结果显示,花生LACS1基因全长2 219 bp,包含1 992 bp的ORF,编码663个氨基酸,有22个外显子和21个内含子。氨基酸序列比对显示花生LACS1有真核生物酰基辅酶A合成酶保守结构域,并含有保守的激活位点和绑定位点。同源性分析发现花生LACS1与大豆、野生大豆、鹰嘴豆、绿豆、甜橙等15种物种的氨基酸序列同源性在68%~86%之间,进化树分析显示,花生LACS1与鹰嘴豆等豆科植物亲缘较近。实时荧光PCR分析表明,花生LACS1在花生根、茎、叶、针、仁和花等组织均有表达,但差异明显,其中花的表达量最高,表达量大小顺序为花>针>叶>茎>根>仁,地上组织表达量高于地下组织。花生LACS1可能参与花生角质层的脂质合成。本研究结果为揭示植物脂肪酸代谢提供理论依据。
Long chain acyl-Co A synthetase (LACS) is an important catalytic enzyme for lipid metabolism. In this study, we cloned the gene sequence from Arachis hypogaea L. to LACS1 (Gen Bank accession number: KT932703) by RT-PCR. The structure of the gene was analyzed, and the homology of the encoded amino acids with other plants was predicted. Real-time Real Tissue expression of LACS1 was studied by -Time PCR technique. The results showed that the LACS1 gene of peanut was 2 219 bp in length and contained a 1 992 bp ORF encoding 663 amino acids with 22 exons and 21 introns. Amino acid sequence alignment showed that peanut LACS1 has a conserved acyl-CoA synthetase domain containing conserved activation sites and binding sites. Homology analysis showed that amino acid sequence homology between peanut LACS1 and 15 species of soybean, wild soybean, chickpea, mung bean and sweet orange ranged from 68% to 86%. Phylogenetic tree analysis showed that the homology of LACS1 between peanut LACS1 and olecranon Beans and other leguminous close relatives. Real-time PCR analysis showed that peanut LACS1 was expressed in the roots, stems, leaves, needles, kernels and flowers of peanut, but the difference was obvious. Among them, the expression of flowers was the highest, the order of expression was flower> needle> leaf> stem> Root> benevolence, aboveground tissue expression was higher than underground tissue. Peanut LACS1 may be involved in the lipid synthesis of peanut stratum corneum. The results of this study provide a theoretical basis for revealing plant fatty acid metabolism.