大鼠神经元Arnt2基因cDNA序列的分析  

Analysis of cDNA sequence of gene Arnt2 from rat cerebellar granule neurons

作　　者：孙林光[1] 银巍[2] 黄奕俊 苏兴文[1] 邱鹏新[1] 颜光美[1] 

机构地区：[1]中山大学基础医学院药理教研室 [2]中山大学基础医学院生物化学教研室,广东广州510089

出　　处：《中国现代医学杂志》2006年第7期974-977,共4页China Journal of Modern Medicine

基　　金：国家自然科学基金项目(No.30472010);广东省自然科学基金团队项目(No.039191)

摘　　要：目的克隆并分析神经元凋亡差异表达5号EST全长cDNA。方法以大鼠大脑MarathonReadycDNA为模板,采用SMARTRACE技术克隆神经元凋亡差异表达5号EST全长cDNA,产物亚克隆到pGEM-Teasy质粒载体后进行序列测定以及同源性分析。结果经过3次5′RACE扩增,得到的5号ESTcDNA长度为5663bp,其中包含1个2136bp的完整的开放读码框序列,与已知大鼠基因芳香烃受体核转位因子(2Arylhydrocarbonreceptornucleartranslocator2,Arnt2)完全同源;该cDNA3′端非编码区长达3323bp,包含3个AATAAA加尾信号和1个由12个腺嘌呤核苷酸组成的polyA序列。结论5号EST目标基因为大鼠基因Arnt2,该基因3′端完整的非编码区cDNA序列为首次报道。神经元凋亡差异表达基因Arnt2的克隆鉴定为深入研究小脑颗粒神经元低钾性凋亡机制奠定了基础。[Objective] To clone and analyse full-length eDNA of neuron apoptosis differentially expressed EST 5. [Methods] Rat brain Marathon Ready cDNA was used as cDNA template, 5＇ SMART RACE was performed for three times, the products of 5＇ SMART RACE were subcloned into pGEM-T easy vector and sequenced; alignment assay was performed with BLAST in NCBI and BLAT in UCSC. [Results] EST 5 was amplified into a sequence of 5 663 bp which containing an ORF with the length of 2 136 base pairs. It was identified that the ORF is homologous to that of rat aryl hydrocarbon receptor nuclear translocator 2 （Arnt2） completely by BLAST and BLAT assay, and it was also identified that this cDNA sequence contained 3 polyadenylation signals of AATAAA and a polyA tall composed of 12 adenines. [Conclusion] The target gene of EST5 is Arnt2. It is the first time that the integrity 3＇ untranslated region of rat gene Amt2 was reported. Identification of differentially expressed gene Arnt2 will be helpful to elucidate apoptosis mechanism of cerebellar granule neurons evoked by low potassium.

关 键 词：神经元 基因 Arnt2 CDNA 

分 类 号：Q71[生物学—分子生物学] R966[医药卫生—药理学]