丹参晚期胚胎蛋白基因SmLEA的克隆及其表达分析.pdfVIP

生物技术通报 BIO TE CHN OLOG Y B ULLETIN 2009 5 SmLEA 刘丛玲 王喆之 ( , 710062) : 对丹参EST序列进行 Blast分析, 发现一条序列与晚期胚胎丰富蛋白( late embryogenesis abundant)基因有 高 的相似性, 在此基础上设计 引物, 分别从 c NA 和 g NA 水平克隆到该基因的全长 ( Genbank 注册号: AY725206) , 命名为 SmLEA该序列全长 739 bp, 无内含子, 包含 1 个长为 495 bp的开放阅读框, 编码 164 个氨基酸序列比对结果显示, 该序列 与番茄的晚期胚胎丰富蛋白 Lemm i9有 高的相似性( 69% ) ,推测该编码蛋白属于晚期胚胎蛋白 LEA 14家族成员生物信息 学显示, SmLEA 所编码蛋白 SmLEA的相对分子质量为 1734 k , 理论等电点为 451, 富含天冬氨酸及ASIPKVVSTIP肽 段, 定位于细胞质中,为稳定类蛋白实时荧光定量 PCR 结果显示, 该基因在 丹参的根茎叶中均有表达, 为组成型表达 基因 : 晚期胚胎丰富蛋白 生物信息学 组成型基因 丹参 Cloning and Expression Analysis ofSmLEA from Sa lvia m iltiorrh iza Bunge Liu Congling W ang Zhezhi (College of Lfie Sciences, ShaanxiNormal University, K ey Laboratory of Med icinalP lant esources andNatural Pharmaceutical Chem istry, M inistry of Education, NationalEngineering Laboratory f or esourceDevelop ing of Endangered Chinese CrudeDrugs inNorthw est of China, X ian 710062) A bstrac:t By analyzing EST sequences ofSalvia m iltiorrh iza , one sequenceswhich showed high homologyw ith late embryogene sis abundant proteins were foundThe fulllength c NA and NA sequence of the genewere cloned and named as SmLEA (GenBank accession number: AY725206)SmLEA consists of739 nucleotides, including a single 495 bp opening reading frame and encoding 164 am ino acidpeptidewithout introns SmLEA shared high amino acid sequence homology ( 69% ) with Lemmi9, a tomato late embryo genesis abundant protein and could be classified into the LEA14 fam ily according to its sequence characterizationBioinformatics analy sis showed that SmLEA w ith a calculatedmolecular mass of 1734 k and a theoretical isoelectric point of 451, was rich in aspartic and AS