Screening differentially expressed genes involved in apoptosis of primary cultured rat cerebellar granule neurons by mRNA differential display RT-PCR
- VernacularTitle:应用mRNA差异显示技术筛选大鼠小脑颗粒神经元凋亡模型中差异表达基因
- Author:
Wei YIN
;
Yijun HUANG
;
Rongbiao PI
;
Xingwen SU
;
Lingzhi ZHAO
;
Pengxin QIU
;
Guangmei YAN
- Publication Type:Journal Article
- Keywords:
Apoptosis;
Differentially expressed genes;
Cerebellum;
Neurons
- From:
Chinese Journal of Pathophysiology
2000;0(11):-
- CountryChina
- Language:Chinese
-
Abstract:
AIM: To Screen and identify differentially expressed genes that involved in apoptosis model in rat cerebellar granule neurons (CGNs).METHODS: The rat cerebellar granule neurons were isolated and primarily cultured. Fluorescent differential display RT-PCR (FDD RT-PCR) was performed to screen differentially expressed ESTs in the apoptosis model of primarily cultured rat CGNs. ESTs were subcloned into pGEM-T EasyTM vector and then sequenced. Alignment assay in non-redunant database was applied for encoding information. Reverse Northern blotting was used to appraise the results from DDRT-PCR.RESULTS: 164 pieces of differentially expressed ESTs were obtained by FDDRT-PCR. 17 of them were subcloned and sequenced. 5 ESTs of 17 were confirmed to be positive results by reverse Northern blotting. CONCLUSION: DD-PCR is a rapid, simple-operation and sensitive method for screening differentially expressed genes, which would contribute to the molecular mechanisms of apoptosis/survive of CGNs.
