用DD-PCR法克隆AVP_(4～8)诱导的鼠脑差异表达基因  

Cloning of a Gene in Rat Brain Induced to Be Expressed Differently by AVP 4-8 with Differential Display PCR

作　　者：熊缨[1] 王迎[1] 刘小龙[1] 杜雨苍[1] 

机构地区：[1]中国科学院上海生物化学研究所,上海200031

出　　处：《生物化学与生物物理学报》1999年第5期509-512,共4页

基　　金：国家攀登计划;国家自然科学基金;上海市科技发展基金

摘　　要：精氨酸加压素（ Ａ Ｖ Ｐ） Ｃ 端片段 Ａ Ｖ Ｐ４ ～８ ， 具有增强记忆的功能， 它在大鼠脑内引发一系列的生理和生化变化。采用差示 Ｐ Ｃ Ｒ（ Ｄ Ｄ Ｐ Ｃ Ｒ） 技术， 寻找注射 Ａ Ｖ Ｐ４ ～８ 前后在大鼠海马中差异表达的基因。抽提总 Ｒ Ｎ Ａ， 以反转录产生的ｃ Ｄ Ｎ Ａ 为模板进行 Ｐ Ｃ Ｒ， 经过９ 组引物组合， 获得了十几个差异片段， 挑选其中差异最大的片段ｄｄ１ 进行克隆、测序， 并与 Ｇｅｎｅｂａｎｋ 等数据库进行同源比较， 没有找到同源基因， 可能是个新基因。而且， Ｎｏｒｔｈｅｒｎ 杂交的结果表明ｄｄ１ 是 Ａ Ｖ Ｐ４ ～８ 刺激产生的差异基因。As a metabolite of arginine vasopressin, AVP 4—8 has been shown to have potent memory enhancing activity and induce a series of physiological and biochemical events in rat brain. The technique known as differential display polymerase chain reaction(DD PCR)was applied to explore genes induced to be expressed differ ently by AVP 4—8 in rat hippocampus. Nine different primer pairs were used to perform DD PCR after reverse transcription, and more than ten differential display fragments were observed. The most remarkable different fragment, dd1, was further cloned and sequenced. Homology searching in Genebank showed that dd1 may be a new gene. Moreover, the results from Northern blot confirmed that dd1 is indeed a gene induced by AVP 4—8 .

关 键 词：克隆 精氨酸加压素 差示PCR 脑 

分 类 号：Q78[生物学—分子生物学] Q426