高、低正加速度耐力大鼠脑组织差异表达基因的分离  

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

作　　者：王晓丽[1] 刘丽[2] 郭丽[1] 范洪学[1] 

机构地区：[1]吉林大学共公卫生学院毒理学教研室,吉林长春130021 [2]解放军空军总医院空军分子生物学中心实验室

出　　处：《吉林大学学报（医学版）》2002年第5期449-453,共5页Journal of Jilin University：Medicine Edition

基　　金：全军"十五"医药卫生基金重点课题 (0 1Z0 16 );空军后勤部"十五"规划项目

摘　　要：目的 :分离高、低正加速度耐力大鼠脑组织差异表达的 c DNA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 m RNA。以高 + Gz耐力大鼠脑组织 m RNA为实验组 ,以低 + Gz耐力大鼠脑组织 m RNA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 c DNA片段混合物 ,然后将此混合物与 p GEM- T easy载体连接 ,转化 E.coli JM1 0 9感受态细胞。结果 :初步获得了 60 8个可能在高 + Gz耐力大鼠脑组织中上调表达的 c DNA片段。这些 c DNA片段长度介于 1 5 0～1 40 0 bp之间 ,集中于 30 0～ 70 0 bp。结论 :在高、低 + Gz耐力大鼠脑组织中差异表达的 c DNA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。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.

关 键 词：正加速度 耐力 大鼠 脑组织 差异表达基因 分离 

分 类 号：Q78[生物学—分子生物学] R852.21[医药卫生—航空、航天与航海医学]