鼠源抑癌新基因Ndr2的克隆、测序与生物信息学分析  

Cloning, Sequencing and Bioinformatics Analysis of a New Tumor Suppressor Gene Ndr2 from Mouse

作　　者：杨兴龙[1] 张亚历[2] 药立波[1] 刘新平[1] 季少平[1] 邢飞跃 

机构地区：[1]第四军医大学生物化学与分子生物学教研室,陕西西安710032 [2]第一军医大学附属南方医院消化病研究所,广东广州510515 [3]第一军医大学全军微循环重点实验室,广东广州510515

出　　处：《癌症》2003年第3期230-234,共5页Chinese Journal of Cancer

基　　金：国家自然科学基金杰出青年基金(编号:39825113)

摘　　要：背景与目的:人源Ndr2(N-mycdown-streamregulator2)基因是本室于1999年从正常人全脑cDNA文库中克隆到的一个新基因犤GenBank,AF159092犦。初步实验表明,Ndr2有可能是一个抑癌基因。为进一步研究该基因的功能,我们拟克隆小鼠的Ndr2的基因组序列。方法:以小鼠的基因组文库为模板,采用RT-PCR方法扩增Ndr2基因的基因组片段,用310GeneticAnalyzer自动测序仪测序,GenBankBLAST相似性分析其与人源Ndr2基因同源性,PcGene和ORFFinder作读框分析,ProDom软件作结构域分析。结果:经琼脂糖DNA电泳鉴定,获得一约3310bp大小的特异条带,并克隆入pMD18-T载体。BLAST相似性分析结果表明该序列与人源Ndr2基因高度同源(同源性为91.4%),与鼠基因组数据库无同源性。已公布的鼠源Ndr2mRNA序列比较发现该基因有8个外显子,7个内含子。读框分析表明,该序列编码含200个氨基酸的蛋白质。ProDom软件分析发现其含有酰基携带蛋白(ACP)样结构域。结论:本研究克隆了一个鼠的Ndr2基因并已测序。该基因为一新基因,该序列已被GenBank收录(登录号:AY151387)。BACKGROUND &OBJECTIVE: Ndr2 (N myc down stream regulator) gene in human is a new gene cloned with the human adult whole brain cDNA as template in 1999, which accession number is AF159092 in GenBank. Locating backward position of the N myc gene in human chromosome, this gene was named Ndr2 gene. The previous experimental results showed Ndr2 gene probably is a tumor suppressor gene. To research the function of Ndr2 gene, the authors cloned the genomic sequence of ndr2 from mouse. METHODS: To clone Ndr2 genomic sequence by reverse transcription polymerase chain reaction(RT PCR) with the mouse genome library as template; automatic sequencing was performed using 310 Genetic Analyzer; homogeneous analysis was made using GenBank BLAST; open reading fragment(ORF) analysis was made using PC Gene and ORF Finder; domain analysis was made using ProDom system. RESULTS: A fragment (about 3310bp,identified by agarose gel electrophoresis) was obtained using RT PCR with the mouse genome library as template. The fragment was cloned in pMD18 T vector. BLAST analysis showed that the sequence was highly homogeneous(with the homogeneity rate of 91.4%) with Ndr2 gene in human and non homogeneous with genomic sequence database in mouse. ORF analysis showed that there was a complete coding region in it, which including 8 extrons and 7 introns; it can interpret a protein containing about 200 amino acid residuals. ProDom analysis showed there was a domain like acyl carrier protein(ACP) in it. CONCLUSION: The authors cloned Ndr2 gene in mouse and proved that the sequence is a new genome sequence in mouse genomic sequence database. At present, the genome sequence has been submitted to GenBank(the accession number: AY151387).

关 键 词：基因 Ndr2 克隆 生物信息学 

分 类 号：R346[医药卫生—基础医学]