花生长链酰基辅酶A合成酶1基因(LACS1)的克隆、鉴定与组织表达

Cloning, Identification and Tissues Expression of Long Chain Acyl-CoA Synthetase 1 Gene (LACS1) in Peanut (Arachis hypogaea L.)

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 coenzyme A synthetase(LACS) is one of the important catalytic lipid metabolic enzymes. In this study, the LACS1 c DNA was cloned from peanut by RT-PCR technique(Gen Bank accession number: KT932703). The gene structure was analyzed, the homology of predictive coding amino acid was analyzed with other plants, and the expression of LACS1 was studied in different tissues by Real-Time PCR.Results showed that the c DNA of peanut LACS1 was 2 219 bp and included an open reading frame of 1 992 bp encoded 663 amino acids. Peanut LACS1 had 21 exons and 22 introns. The sequence alignment of amino acid has revealed eukaryotic long-chain fatty acid Co A synthetase conserved domain, and contains the conserved activation sites and the binding site. Homology analysis found that peanut LACS1 had homology between 68% and 86% with soybean, wild soybean, chickpeas, mung beans, and sweet orange species. The evolutionary tree indicated peanut LACS1 had close kinship with leguminous plants such as chickpeas. Real-time PCR analysis revealed that peanut LACS1 was differentially expressed in needle, root, stem, leaf, flower and kernel and the flowers was the highest.The expression quantity of overground parts was higher than the underground. The order of expression quantity was flower>leaf>stem>root>needle>kernel. Peanut LACS1 may be involved in the synthesis of peanut cuticle. The results provided theoretical basis for the further research of plant fatty acid metabolism and quality improvement of plant.