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树鼩PSEN1全长编码序列的克隆及分子特征分析 预览

Cloning and characterization of presenilin 1 in the Chinese tree shrew
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摘要 目的 获取树鼩早老素蛋白-1(PSEN1)的全长编码序列并进行分子特征分析。方法 以树鼩脑组织总RNA为材料,通过RT-PCR、RACE-PCR扩增和序列拼接获得PSEN1基因全长编码序列,进而通过DNAMAN、MEGA等生物信息学软件对其序列和分子特征进行分析。qRT-PCR和Western Blot进一步分析PSEN1在树鼩各个组织的表达模式。结果 克隆鉴定了树鼩PSEN1基因,其cDNA的开放阅读框全长1128 bp,编码375个氨基酸。通过系统发育谱系树、氨基酸序列对比分析,发现树鼩PSEN1与小鼠、大鼠等相比更接近人类和非人灵长类动物。qRT-PCR和Western Blot的结果表明,树鼩PSEN1在脑组织的表达量明显高于其他脏器组织。结论 通过克隆树鼩PSEN1基因序列并进行分析,为今后进一步深入研究该基因功能和建立相关疾病动物模型提供理论基础。 Objective To get the full-length presenilin-1(PSEN1)cDNA encoding sequence and analyze its molecular characteristics.Methods Full-length PSEN1 cDNA was cloned from total RNA from brain tissue of tree shrew by RT-PCR and RACE-PCR.The molecular characteristics were compared with PSEN1 in other mammals and evaluated using biology softwares such as DNAMAN,MEGA and others.Quantitative reverse transcription PCR and western blot assays were used to examine the mRNA and protein expression pattern of PSEN1 in various tissues of tree shrew.Results The open reading frame sequence of PSEN1 was 1128 bp and encoded 375 amino acids.We constructed a phylogenetic family tree and compared their PSEN1 amino acid sequences.Tree shrew PSEN1 was closer to humans and non-human primates than mouse and rat.The expression of PSEN1 mRNA and protein was obviously higher in the brain than that of other organ tissues.Conclusions We successfully cloned the PSEN1 gene in tree shrew and provided comparative analysis with other species.These findings provide a theoretical basis for further study on the function of this gene and the establishment of animal models of Alzheimer’s disease(AD).
作者 李明学 王文广 李娜 匡德宣 仝品芬 黄鑫 黎晓慧 孙晓梅 陆彩霞 LI Mingxue;WANG Wenguang;LI Na;KUANG Dexuan;TONG Pinfen;HUANG Xin;LI Xiaohui;SUN Xiaomei;LU Caixia(Tree Shrew Germplasm Resource Center,Institute of Medical Biology,Chinese Academy of Medical Sciences (CAMS) & Peking Union Medical College(PUMC),Kunming 650118,China;Kunming Medical University,Kunming 650500)
出处 《中国实验动物学报》 CAS CSCD 北大核心 2019年第4期456-465,共10页 Acta Laboratorium Animalis Scientia Sinica
基金 国家自然科学基金(31601907) 云南省科技人才和平台计划项目(2017HC019、2018HB071) 云南省重大科技专项(2017ZF007)。
关键词 树鼩 PSEN1 分子克隆 系统发育分析 tree shrew PSEN1 cloning phylogenetic analysis
作者简介 李明学(1995—),男,在读硕士研究生,研究方向:实验动物学。Email:18687520819@163.com;通信作者:孙晓梅(1963-),女,主任技师,研究生导师。Email:sxm@imbcams.com.cn;通信作者:陆彩霞(1979-),女,博士,研究生导师。Email:lcx@imbcams.com.cn.
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