摘要
目的克隆人乳腺癌ＭＣＦ－７细胞凋亡相关基因，分析验证所用方法的特点。方法采用基于ＰＣＲ的消减杂交技术，在已建立的全反式维甲酸诱导人乳腺癌ＭＣＦ－７细胞的凋亡模型中，克隆ＭＣＦ－７细胞凋亡的相关基因。结果从１３个克隆中，共筛选出５个表达的基因。其中４个为已知基因，１个为新基因。新基因命名为ａｐｍｃｆ－１，Ｇｅｎｂａｎｋ登录号为ＡＦ１４１８８２。４个已知基因中３个与凋亡关系密切。结论这种改良的基于ＰＣＲ的消减杂交技术，为克隆差异表达基因提供了一种新的方法和思路。
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 deferential expression gene.
出处
《细胞与分子免疫学杂志》
CAS
CSCD
北大核心
2001年第1期35-37,共3页
Chinese Journal of Cellular and Molecular Immunology
