Improved PCR-based subtractive hybridization, a new strategy on cloning differential expression genes in apoptotic MCF-7 cells
10.3321/j.issn:1007-8738.2001.01.010
- VernacularTitle:克隆 MCF-7细胞凋亡差异表达基因的一种方法
- Author:
Wei YAN
;
Feng ZHU
;
Zhongliang ZHAO
;
Yubo CAI
;
Wen YUE
;
Chen SHAO
;
Fan LU
;
Qing LI
;
Chengji WANG
- Publication Type:Journal Article
- From:
Journal of Cellular and Molecular Immunology
2001;17(1):35-37
- CountryChina
- Language:Chinese
-
Abstract:
Aim To clone apoptosis-related genes from human MCF-7 breast cancer cells and to analyze the character of the method used in the process. Methods A poptotic cell model of MCF-7 cells was established with the apoptotic tumor cells induced by the all-trans-retinoic acid. The apoptotic gene was cloned from the model by improved PCR-based subtractive hybridization. Results 5 clones were identified to be related to apoptosis by reverse dot blot, 4 of them were known genes, and 3 were related to apoptosis. A novel gene， named apmcf-1, coded for 47 amino acid was identified. This gene was accepted by Genbank, the accession number was AF141882. Conclusion This improved PCR-based subtractive hybridization may be an efficient way in cloning differential expression gene.
