八肋游仆虫一种富含三核苷酸重复序列的新基因GARP的克隆和序列分析.pdfVIP

HEREDITAS (Beijing) 2007 1 , 29(1): 87―91 ISSN 0253-9772 研究报告 DOI: 10.1360/yc-007-0087 GARP 许静, 王伟, 柴宝峰, 梁爱华 , , 030006 : 人类基因中三核苷酸重复序列拷贝数的异常扩增, 可导致多种神经系统疾病。一种富含GAA 三核苷酸的 GARP (glutamic acid-rich protein)基因从八肋游仆虫(Euplotes octocarinatus) 大核文库中筛选获得。大核中该基因 的染色体全长460 bp, 基因两端具有下毛类纤毛虫大核特有的端粒序列(C A C A C A C ), 开放读框内含有一 4 4 4 4 4 4 4 个TGA(88-90) 密码子, 在游仆虫中编码为半胱氨酸。经DNA Star 软件分析, 该基因编码的蛋白质由 112 个氨基酸 组成, 预测其分子量为 13 kDa, 等电点为3.82, 含有四个 α 螺旋和一个β 折叠。小核中对应的该基因含有两个 内部删除序列, IES1 和IES2 。IES1 和IES2 分别长41 bp, IES1 以GA 二核苷酸直接重复为删除信号, IES2 以TA 二核苷酸直接重复为删除信号。RT- PCR 证明该基因具有转录活性。 : 八肋游仆虫; GARP 基因; 三核苷酸重复 Cloning and characterization of a novel trinucleotide repeat- containing gene GARP from Euplotes octocarinatus XU Jing, WANG Wei, CHAI Bao-Feng, LIANG Ai-Hua Key Laboratory of Chemical Biology and Molecular Engineering of Ministry of Education, Institute of Biotechnology, Shanxi University, Taiyuan 030006, China Abstract: The expansion of trinucleotide repeats in genome is related to the phthogenesis of several neurodegenerative deseases. A GARP (glutamic acid-rich protein) gene was isolated from the macronuclear plasmid mini library of Euplotes octocarinatus. A micronuclear version of the GARP gene was amplified by polymerase chain reaction. The macronuclear DNA molecule carrying the GARP gene is 460 bp long and shows the characteristics of macronuclear chromosomes of hy- potrichous ciliates. One of the three cysteines is encoded by the opal codon TGA(88-90). The predicted open reading frame encodes a 112-amino acid polypeptide, with a predicted molecular mass of 13 kDa and an isoelectric point of 3.82. Mic