用差异显示法从人胎脑基因文库分离一个编码序列  

Separation of An Encoded Sequence from Human Fetal Brain Library by mRNA Differential Display

作　　者：杨岐生[1] 林卿[1] 李奕[1] 谢毅[2] 

机构地区：[1]浙江大学生物科学与技术系,杭州310027 [2]复旦大学遗传学国家重点实验室,上海200433

出　　处：《中国生物化学与分子生物学报》1999年第6期876-880,共5页Chinese Journal of Biochemistry and Molecular Biology

基　　金：复旦大学遗传学国家重点实验室开放项目

摘　　要：人１８周、２２周胎儿脑、肝肾组织总ｍ ＲＮＡ 用ＤＤＲＴ－ＰＣＲ显示出差异的条带，回收胎脑和肝肾特异性表达的４８７条电泳条带．其中某些条带用３种组织的ｃＤＮＡ 探针作点杂交，筛选只对胎儿脑总呈阳性的ＤＮＡ 片段．以其中某一条带ＤＮＡ 为探针，从胎儿脑ｃＤＮＡ 文库筛选阳性克隆，得到ＧＣ５８．经Ｎｏｒｔｈｅｒｎ 杂交和ＤＮＡ 测序，表明它是人脑表达的序列，与数据库中ＫＩＡＡ０５１５有同源性，并编码一个有２７４个氨基酸的蛋白质，该蛋白质序列尚未见报道．探讨了ＤＤＲＴ－ＰＣＲ的条件和假阳性问题．DDRT PCR of total mPNA from human fetal brain or liver and kindey showed different bands.487 bands specific for these tissues were recovered.Some of them were hybridized by dot blot with 3 kinds of cDNA probes from brain or liver and kindey mRNA to screen brain specific bands.A clone,GC58,was obtained from fetal brain cDNA bank using a brain specific band DNA as probe,and identified in Northern hybridization.GC58 DNA sequence showed that it was expressed in human brain and was homologous to KIAA0515 sequence.The read frame encodes a 274 amino acid peptide which has not been reported.

关 键 词：胎儿 人脑 DDRT-PCR DNA序列 基因分离 假阳性 

分 类 号：Q781[生物学—分子生物学] Q－33