银染DD-PCR法筛选游泳疲劳小鼠肝脏中差异表达基因的初步研究  

STUDY ON SCREENING DIFFERENTIALLY EXPRESSED GENES IN MICE LIVERS BY SILVER STAINING DD-PCR

作　　者：栾新红[1] 胡仲明[1] 刘维全[2] 江禹[1] 柳巨雄[1] 王凯[1] 

机构地区：[1]军事医学科学院军事兽医研究所 [2]中国农业大学生命科学院,北京110092

出　　处：《中国应用生理学杂志》2005年第3期296-299,共4页Chinese Journal of Applied Physiology

基　　金：总后勤部课题(需030128)

摘　　要：目的:筛选小鼠游泳疲劳相关基因,为阐明疲劳产生的分子机制奠定基础。方法:取30只BALB/c雄性小鼠,体重(20±2)g,随机分成对照组、浸水组和游泳疲劳组,对游泳疲劳组小鼠进行负重游泳致疲劳后,与其他两组同时采集肝脏组织,利用改进后的银染DD-PCR法筛选小鼠肝脏中差异表达的基因,并进行鉴定,采用BLAST软件对阳性片段进行同源性分析。结果:经反向Northernblot和测序鉴定后,获得了7条阳性差异表达基因片段(DD-ESTs),其中有2条DD-ESTs只在游泳疲劳组表达,2条呈现下调表达,另3条呈现上调表达;阳性片段中一条为新基因,登录GenBank数据库后获得登录号为AY615302。结论:利用银染DD-PCR法获得了7条游泳疲劳小鼠差异表达基因片段(DD-ESTs)。Aim： To screen swimming-fatigue related genes in mice and lay theoretic basis for researching the molecular mechanism of fatigue. Methods： 30 male BALB/c mice（20 ± 2g） were divided into control group, dipping in water group and swimming-fatigue group respectively. After fatigue for swimming in swimming-fatigue group, with control group and dipping in water group, liver tissues in mice were collected. With improved silver staining mRNA differential display method, the differentially expressed genes in mice livers were screened and evaluated by reversed Northern blot. The positive segments were analyzed homology by BLAST. Resuits： 7 of DD-ESTs were gained. Two of them only expressed in swimming-fatigue group, two down-regulated expressed, and three up-regulated. One of them was a novel gene and was accepted by GenBank, AY615302. Conclusion： Seven DD-ESTs in swimmingfatigue mice were gained by silver staining mRNA differential display method.

关 键 词：疲劳 DD—PCR法 肝脏 小鼠 

分 类 号：Q78[生物学—分子生物学] S852.2[农业科学—基础兽医学]