中国医学科学杂志(英文版)
中國醫學科學雜誌(英文版)
중국의학과학잡지(영문판)
CHINESE MEDICAL SCIENCES JOURNAL
2000年
4期
195-200
,共6页
金虎林%胡松年%李光涛%涂纯%袁建刚%强伯勤
金虎林%鬍鬆年%李光濤%塗純%袁建剛%彊伯勤
금호림%호송년%리광도%도순%원건강%강백근
glioma%all-trans retinoic acid%differential display reverse transcribed-PCR( DDRTPCR)
Objective. To investigate the differentiation process of the human glioblastoma cells.
Methods. Differential display reverse transcribed-PCR(DDRT-PCR) was used to isolate the genes differentially
expressed in control and all-trans retinoic acid treated human glioblastoma cell line BT-325. Routine method of cDNA
library screening was performed to clone full-length cDNA.
Results. Thirty-six RT-PCR reactions were performed and 64 differentially expressed fragments were recovered,
amplified and cloned. Of them,46 ESTs were sequenced and delivered into the GenBank. The homology comparison us
ing BLAST algorithm revealed that 22ESTs are highly homologous with the known genes and many of them play impor
tant roles in the cell differentiation progress. A dot-blot hybridization was conducted to certify the differentiation expres
sion. The result showed that 27 EST clones are expressed at different level in control and all-trans retinoi c acid treated
BT-325 cells. A full-length cDNA was cloned using the ES T-HGBB098.
Conclusion. DDRT-PCR was a simple and effective method to serially analyze the differentially expressed genes.