Identification of some expressed sequence tags in cerebellum of rat / 中国病理生理杂志

ABSTRACT

AIM: To obtain differentially expressed cDNA fragments in the cerebellum of rats and screen unknown expressed sequence tag (EST). METHODS: Suppression subtractive hybridization (SSH) was carried out, in which the cDNA fragments of cerebellum were taken as "tester" and correspondingly that of cerebrum and brain stem as "driver". The homogeneous sequences between the tester and driver were excluded and the rare sequences in cerebellum were enriched by SSH. The differentially expressed cDNA fragments were further cloned for the construction of subtracted cDNA libraries and sequencing. RESULTS: 32 clones were selected and 34 cDNA fragments were sequenced. 8 of 32 were proved to be true positive with reverse Northern assay, 13 of 34 fragments were identified to be new cDNA fragment and given the gene sequence numbers by GenBank (AW288461-AW288474). CONCLUSION: SSH is very useful method for screening differentially expressed genes. Our data may be helpful to understand the molecular mechanism of brain function.