小鼠Doc-1R基因的克隆及其表达  

Cloning and Expression of the Mouse Doc-1R Gene

作　　者：生秀杰[1] 周伟强[2] 姜莉[2] 张梅英[1] 王太一[1] 张学[2] 

机构地区：[1]中国医科大学实验动物部遗传室 [2]中国医科大学卫生部细胞生物学重点实验室,辽宁沈阳110001

出　　处：《癌症》2002年第2期122-122,共1页Chinese Journal of Cancer

基　　金：国家"九五"攻关项目(96-A230602);国家自然科学基金(39980011)

摘　　要：背景与目的:Doc-1R基因是1999年克隆的一种新的基因,已有的研究表明Doc-1R基因可能是一种潜在的抑癌基因。为了进一步研究此基因的功能,我们首先克隆了小鼠的Doc-1R基因,并对此基因的表达进行了初步的研究。方法:根据小鼠的Doc-1R基因cDNA序列,设计合成基因组特异引物,应用巢式PCR对小鼠基因组步移文库进行扩增。对测序结果进行序列分析及剪接位点的鉴定。另外应用RT-PCR的方法研究了Doc-1R基因在小鼠肝、脾、胰、肾、肺、肠、心、脑、骨、肌肉、膀胱、卵巢、睾丸等13种组织的表达。结果:经二次基因组步移获得了小鼠Doc-1R基因组全长序列。基因组全长2787bp,含4个外显子和3个内含子,外显子与内含子接头符合GT/AG法则。RT-PCR结果表明,Doc-1R基因在小鼠13种组织和器官中均有表达。结论:成功克隆了小鼠Doc-1R基因,为进一步研究此基因的功能奠定了基础。RT-PCR表达结果提示此基因可能是对维持组织器官的功能具有重要的作用的管家基因。Background &Objective:Doc-1R gene is a new gene which was clo ned in 1999.Recent studies suggest that Doc-1R gene is a potential tumor suppressor gene.In order to study th e function of the Doc-1R gene,The aim of this study was to obtain its genomic sequences and to analyze its expression in the tissues.Method:According to the cDNA sequences of Doc-1R gene,the gene-s pecific primers were designed and sy nthesized.The sequence of Doc-1R ge ne was cloned through nested PCR using the Genome Walker kit.The sequence and sp lice donor /acceptor site were analy zed.Using the reverse transcription polymera se chain reaction(RT-PCR),the expression of Doc-1R gene in thi rteen tissues,including mouse liver,spleen,pancreas,kidney,lung,intestine,heart,brain,b one,muscle,bladder,ovary,sperma ry were determined.Results:The mouse Doc-1R gene has been obtain ed by two times genomic walking.This gene spans 2787bp and contains four exons and three int rons.All of the splice donor /accept or site sequences were in accordance with the consensus'GT-AG'rule.RT-PCR experiments demonstra ted Doc-1R gene was expressed in the 13tissue samples.Conclusion:The authors have successfully cloned the mouse Doc-1R gene,which will be the foundation for further investigating the function of that g ene.The expression pattern suggests that the Doc-1R gene is a hou sekeeping gene which is important to keep the function of tis sues and organs.

关 键 词：DOC-1R 基因克隆 DNA序列分析 基因表达 小鼠 癌变 

分 类 号：R730.2[医药卫生—肿瘤]