正加速度重复暴露大鼠脑差异表达基因的筛选

目的 筛选大鼠经正加速度(Gosiliveacceleration, Gz)重复暴露后脑的差异表达基因,探讨 Gz重复暴露致脑损伤的分子机制。方法 应用抑制性消减杂交技术,构建高消减效率的 Gz重复暴露大鼠脑cDNA消减文库;用差异筛选技术筛选阳性克隆;用序列分析确定阳性克隆的性质;用RT—PCR证实阳性克隆的可靠性。结果 从消减库中获得70个阳性克隆;经差异筛选后,选取其中10个阳性克隆,序列分析表明,7个克隆与已知基因高度同源,3个为新的候选基因。RT—PCR证实了差异表达基因在 Gz重复暴露大鼠脑中高表达。结论 用抑制性消减杂交技术筛选发现的3个新的cDNA可能在 Gz脑损伤的病理过程中起重要作用。

Screening of differentially expressed genes in rat brain repeatedly exposed to +Gz

Objective To screen the differentially expressed genes in rat brain repeatedly exposed to +Gz and to explore the possible molecular mechanism of brain damage due to repeated +Gz exposures Methods A cDNA subtractive library with high subtractive efficiency of repeated +Gz exposures in rat brain was constructed by suppression subtractive hybridization (SSH) Positive clones were screened by differential screen technique Sequence analysis was performed on 10 clones The differentially expressed genes were confirmed by RT PCR Results Seventy positive clones were identified from SSH pools Sequence analysis performed on the 10 positive clones showed that 7 clones were homologous with the genes published in GenBank and 3 clones were unknown genes The results from RT PCR confirmed that the expression levels of those differentially expressed genes in rat brain of +Gz exposure group were significantly higher than those in control group Conclusion The three novel differentially expressed cDNAs are found with SSH and they might play an important role in the pathologic course of rat brain damage due to repeated +Gz exposures