ABSTRACT
Objective To screen the differentially expressed genes in optic tecta of pigeon with monooccular deprivation. Methods Using suppression subtractive hybridization technique,the differentially expressed cDNA library for the left optic tecta of pigeons was constructed;recombinant clones were sequenced randomly and further proved to be the true positive by reverse Northern blot analysis. Results 150 of EST (expressed sequence tags) were sequenced and 16 of them proved true positive,which included those homologous to known genes and those having no matches.138 of ESTs were assigned Accession Number by Genbank.Conclusion\ Our data provided to be the useful clues for studying the development of visual system and the molecular construction of vision center system.
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.