新生大鼠大脑特异表达基因的初步筛选

目的 获得新生大鼠大脑特异表达的cDNA序列,筛选与新生大鼠大脑发育相关的未知的表达序列标签（expressed sequence tag,EST）。方法 以新生大鼠的大脑mＲＮＡ逆转录cDNA作为实验组（tester）,成年大鼠大脑mRNA逆转录cDNA作为对照组（driver）,经抑制消减杂交（suppression subtractive hybridization,SSH)方法消除同成年大鼠大脑相同的cDNA,并富集低丰度基因,克隆建立新生大鼠大脑特异表达的cDNA文库。结果 获得11个cDNA基因片段,同时将测序结果与Genebank进行同源性比较,发现5个首次在大鼠大脑中检测到的基因序列。结论 SSH是目前寻找差异表达基因的最为简便有效的方法,为寻找大鼠大脑发育差异表达基因提供了一些线索。

Screening of Differentially Expressed Genes in Cerebrum of 1 Day Postnatal Rat

Objective To obtain differentially expressed genes in cerebrum of 1 day postnatal rat and to screen unknown expressed sequence tag(EST) relating to the developmental cerebrum of 1 day postnatal rats.Methods Suppression substractive hybridization(SSH) was carried out, in which cDNA fragments from cerebrum of 1 day postnatal rat were taken as "tester" and it corresponded to those of the adult rats as "driver". The homogeneous sequences between the tester and the driver were excluded and the low rebundance was enriched. After cloning ,a differential cDNA library of cerebrum of 1 day postnatal rat was constructed.Results 11 cDNA fragments were obtained. The homogeneity of 11 sequences was compared with Genebank and 5 of 11 cDNA fragments were identified to be novel.Conclusion SSH is the most simple and effective method for seeking differentially expressed genes at present