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本研究利用cDNA-RDA(cDNA Representational DifferencesAnalysis)技术研究了在人红白血病细胞株TF-1细胞撤除细胞因子后进入凋亡时诱导表达的基因.我们以去细胞因子8小时后的TF-1细胞为tester,以正常培养的TF-1细胞为diver,进行三轮RDA.得到的RDA产物克隆至测序载体中进行测序.测序20个克隆经向GeneBank查询发现了6个TF-1细胞凋亡表达或高表达的新基因片段.其中有三个经与GenBank nr和dbest查询均没有发现同源性,已经GenBank进行登记,登记号分别为U83208.U83279,U83397.同时还发现一批已知基因与凋亡相关,其中包括Hou和人疏氧还原蛋白等,以前的工作提示它们在凋亡中起作用.对20个克隆中的13个用Slot Blot进行检测发现其中7个只在去细胞因子的TF-1细胞中表达或高表达,Northem Blot结果与Slot Blot结果一致并估计了两个未知基因的大小约为1.5～2.0kb.本研究为进一步研究凋亡相关基因打下了良好的基础.目前,我们除了进一步研究一些已
In this study, cDNA-RDA (cDNA Representational DifferencesAnalysis) technology to study the human erythroleukemia cell line TF-1 cells after removal of cytokines induced apoptosis expression of genes.We take cytokines in TF-1 cells after 8 hours As a tester, three rounds of RDA were performed on normal cultured TF-1 cells, and the obtained RDA product was cloned into a sequencing vector for sequencing.Sectioning 20 clones, we found 6 TF-1 cell apoptosis genes Or high expression of the new gene fragments.Among three of the GenBank nr and dbest query did not find homology, has been registered GenBank accession numbers were U83208.U83279, U83397.At the same time also found a group of known genes and withered These results suggest that they play a role in apoptosis.Study on 13 out of 20 clones by Slot Blot revealed that only 7 of them were cytokines TF -1 cells, and the result of Northem Blot was consistent with that of Slot Blot and the size of two unknown genes was estimated to be about 1.5-2.0 kb.This study laid a good foundation for further study of apoptosis-related genes At the moment, we have to do some further research