高、低正加速度耐力大鼠脑组织差异表达基因的分离

目的:分离高、低正加速度耐力大鼠脑组织差异表达的cDNA片段,以获得耐力相关基因.方法:雄性SD大鼠在离心机上处理后,选取耐受终点在高、低两个极端的动物,立即取全脑,分离mRNA.以高+Gz耐力大鼠脑组织mRNA为实验组,以低+Gz耐力大鼠脑组织mRNA为驱动组,采用抑制消减杂交技术分离高、低+Gz耐力大鼠脑组织中差异表达的cDNA片段混合物,然后将此混合物与pGEM-T easy载体连接,转化E.coli JM109感受态细胞.结果:初步获得了608个可能在高+Gz耐力大鼠脑组织中上调表达的cDNA片段.这些cDNA片段长度介于150～1 400 bp之间,集中于300～700 bp.结论:在高、低+Gz耐力大鼠脑组织中差异表达的cDNA片段所代表的基因为+Gz耐力相关基因,这些基因的上调表达可能与+Gz耐力的提高有关.

Isolation of differentially expressed genes from rat brains with high and low tolerance to +Gz

Objective:To obtain genes related to tolerance to +Gz through isolation of cDNA fragments that differentially expressed between brains of rat with high and low tolerance to +Gz. Methods: Brians of rats with high and low tolerance to +Gz were isolated immediately and poly A+ RNA of those were extracted after male SD rats were treated on animal centrifuge. cDNA fragments that differentially expressed between brains of rats with high and low tolerance to +Gz were separated using suppression subtraction hybridization with mRNAs of brains of rat with high tolerance to +Gz as tester and those with low tolerance to +Gz as driver,the cDNAs were ligated with pGEM T vector,and then the ligated products were transformed into competent cells of JM109.Results:Six hundreds and eight cDNA fragments that were probably up regulated expression in brains of rats with high tolerance to +Gz were obtained and the length of fragments was ranged from 150 to 1 400 bp.Conclusion:Genes which were differentially expressed between brains of rats with high and low tolerance to +Gz are probably related to tolerance to +Gz with up regulated and may contribute to the improvment of tolerance to +Gz.