人胎脑mRNA的差异显示分析及脑表达新ESTs的分离

应用mRNA差异显示技术分析人胎脑组织不同发育阶段及不同部位基因表达的差异并研究其特异表达的基因。以3个锚定引物Oligo（dT）12M（M=A，G，C）及20个随机引物，对脑组织mRNA进行了近50次差异显示分析，共分离出上百个差示产物。对其中部分产物进行克隆、测序获得了39个CDNA序列，长度为100～500bp。其中34个已被Genbank接受并给予新序列编号。

Differential Display Analysis of Human Fetal Brain mRNAs and Isolation of Brain-specific Novel ESTs

Using the differential display technique(DDRT-PCR) originally developed by Liang andPardee, we tried to isolate the stage-specific or tissue-specific genes in the developing humanneural system (brain). With about fifty sets of arbitrary primers and anchored primers,DDRT-PCR was carried out to analyze the differentially expressed mRNAs between humanfetal brains of different developmental stage or different parts of human fetal brain at thesame developmental stage. About one hundred bands containing cDNA fragments of differ-ential interest were cut out from the polyacrylamide gel, thirty-four of which were clonedand sequenced. They were accepted as novel cDNA sequences by GeneBank. In order tocheck the feasibility of DDRT-PCR,three of the thirty-four cDNA sequences were randomlychosen as probes for further analysis by dot blot RNA hybridization, and two proved to bespecifically expressed in human brain-