【作者】 刘月环； 吴旧生； 石巧娟； 郭红刚； 卢领群； 萨晓婴； 柯贤福； 毛栋森；

【Author】 Liu Yue-huan~1,Wu Jiu-sheng~2,Shi Qiao-juan~1,Guo Hong-gang~1,Lu Ling-qun~1,SaXiao-ying~1,Ke Xian-fu~1, Mao Dong-sen~1 (1 Zhejiang Academy of Medical Sciences,Hangzhou,310013,China) (2 College of animal sciences,Zhejiang university,310029,China)

【机构】 浙江省医学科学院； 浙江大学动物科学学院；

【摘要】 目的克隆长爪沙鼠载脂蛋白E基因(ApoE)的内含子,并对前期克隆到的该基因的编码区进行序列分析与结构初步预测。方法根据Genbank中的相关序列,在前期获得的编码区序列的基础上对基因组进行套式扩增,获得了ApoE基因编码区全长,然后通过对编码区序列进行碱基分析比对后获得2个内含子。同时用RNA Structure5.1软件对该基因mRNA二级结构进行预测。结果长爪沙鼠载脂蛋白ApoE基因包含两个内含子,长度分别为521bp和421bp,经blsat比较发现测序的结果与大鼠ApoE基因(编号为J02582.1)的编码区内含子的同源性达到了80.9%、76.8%,与小鼠ApoE基因(编号为NC 000073.5)的编码区内含子同源性达77.0%、75.1%,与人ApoE基因(NC 000019.9)的同源性达到了48.2%、46.5%,已作为前面所获cds区的补充向genbank提交,登录号为HM366607。mRNA分析表明5’不译区为65bp,3’不译区为119bp,ORF为951bp,茎552bp,环501bp,自由能为-531.0。结论根据比较基因组学原理,用套式PCR可以得到长爪沙鼠载脂蛋白E基因编码区的内含子序列,本研究为筛选长爪沙鼠高脂血症模型的遗传多样性标记打下基础。更多还原

【Abstract】 Objective To clone the apolipoprotein E gene introns in complete cds of the Mongolian gerbil.Analysis and Predict the mRNA motif based on the previous results in the CDS of Apolipoprotein E（APOE） gene.Methods Eight pairs of primers were designed according to the published sequences the relate introns and our previous sequences submitted to GenBank.Nest-PCR was applied to clone the apolipoprotein E introns in liver tissue of Mongolian gerbil.The PCR products was sequenced and blasted with related sequences.At the same times,the mRNA motif of ApoE gene was analysised and preidicted.Results The results of PCR and 421bp respectively in the Mongolian gerbil.The results of blast showed the identification between Mongolian gerbil and rat（J02582.1）is 80.9%、76.8%,the identification between Mongolian gerbil and mouse（NC₀00073.5） is 77.0%、75.1%,the identificatin between Mongolian gerbil and human（NC000019.9） is 48.2%and 46.5%,respectively.As a accessory of a GenBank accession number HM366607.the mRNA show 5-UTR is 65bp,3-UTRis 119bp,ORF is 951bp,stem is 552bp,loop is 501bp,free energy is-531.0.Conclusions The Nest-PCR products were the sequence of the apolipoprotein E two introns of Mongolian gerbil.A further study to explore the genetic diversity of apolipoprotein E gene is expected.更多还原