人18周胎儿脑cDNA文库的构建及鉴定  

Construction and analysis of cDNA library of human 18-week-old fetal brain

作　　者：林卿[1] 杨岐生[2] 殷明[3] 金丽娟[4] 肖谷田[5] 叶蕴辉 刘建平[5] 王玮[2] 谢毅[5] 

机构地区：[1]复旦大学遗传学研究所,上海200433 [2]浙江大学 [3]海军医学研究所 [4]北京协和医科大学 [5]复旦大学

出　　处：《复旦学报（自然科学版）》1997年第5期512-516,共5页Journal of Fudan University：Natural Science

基　　金：复旦大学遗传学国家重点实验室开放基金

摘　　要：许多脑基因的特异性表达对人脑的发育、分化有着重要的作用．为了研究这些基因的结构和功能，构建了一个人18周胎儿脑cDNA文库．抽提总mRNA后，经过一系列的酶促反应合成cDNA．分级分离柱除去小片段后克隆到λgt10载体中．转染宿主菌C600hfl后文库包装效率为4．6×106pfu／μg，cDNA平均插入片段大于1．2kbp．根据已知序列设计引物，从cDNA文库中扩增出神经生长因子（NGF）的全长编码基因．Expression of some genes in fetal brain may take an important role during the development and differentiation of the brain. In order to investigate the function and the structure of these genes. a fetal brain cDNA library was constructed. mRNA was extracted from an 18-week-old human fetal brain. cDNAs were synthesized after a series of enzymatic reac non and then cloned into the vector λgt10 after short fragments to be removed with cDNA size fractionation columns. The packaging efficiency is about 4. 6 ×106 pfu/μg after infectin the host bacteria C600hfl. The average length of the cDNA inserts is about 1. 2 kbp. A fulllength NGF cDNA was amplified from the cDNA library by polymerase chain reacion (PCR) with primers which were designed according to the nucleotide sequence of nerve growth factor (NGF ).

关 键 词：胎儿 人脑cDNA文库 胚胎学 基因 人类遗传学 

分 类 号：R321[医药卫生—人体解剖和组织胚胎学] Q987[医药卫生—基础医学]