关键词: 抑制消减杂交技术；癫痫,颞叶；海马；基因表达

Abstract:

Objective   To construct a subtracted cDNA library of differentially expressed genes in the hippocampus of rats with temporal lobe epilepsy, and to screen for differentially expressed genes related to temporal lobe epilepsy. Methods   Suppression subtractive hybridization (SSH) was used for isolating cDNA fragments of differentially expressed genes in the hippocampus of temporal lobe epilepti rats, and normal hippocampus tissues were controls. The products of SSH were directly inserted into the T/A cloning vector to set up the subtractive library which was amplified through E.coli transformation. One hundred positive bacterium clones were randomly picked and screened the colony polymerase chain reaction (PCR) method.  Results   Fourteen differentially expressed genes were obtained from the hippocampus of temporal lobe epileptic rats by sequencing and using Basic Local Alignment Search Tool (BLAST), and they were highly homologous with the known gene fragments in rats. Conclusion   SSH is an effective method for screening for differentially expressed genes. Many differentially expressed genes exist in the hippocampus of rats with temporal lobe epilepsy. The results of this study provide an important clue for further exploring the molecular mechanism of temporal lobe epilepsy.

Key words: Suppression subtractive hybridization; Epilepsy, temporal lobe; Hippocampus; Gene expression