用mRNA差异显示方法寻找小鼠胸腺基质细胞差异表达基因

Application of DD-PCR to analyse genes expressed in cloned mouse thymus stromal cell lines

目的寻找并初步分析小鼠胸腺基质细胞差异表达基因。方法来源于小鼠胸腺基质细胞系的两株亚克隆细胞 4 ( 5 )和 4 ( 12 ) ,前者可诱导前体 T细胞的进一步成熟 ,分别提取该两株细胞的总 RNA,利用 m RNA差异显示方法( DD- PCR)筛选 4 ( 5 )细胞特异表达的基因片段。结果得到了 17个 4 ( 5 )细胞特异表达的基因片段 ( expressed sequences tag,EST) ,其中 14个代表了尚未登录的小鼠新基因 ,另外 3个则分别与小鼠的 cytochrome C oxidase,Hsp65 ,Eps8基因高度同源。结论 DD- PCR方法是一种优良的寻找新基因的方法 ,小鼠胸腺基质细胞可能分泌或表达更多的新因子或分子。

ObjectiveTo search the genes expressed in cloned mouse thymus stromal cell lines. MethodsImproved DD-PCR was used to detect differentially expressed genes between two cloned mouse thymus stromal cells:4(5) vs 4(12), the 4(5) cells expressed some special protein molecules which can induce progenitor T cells to further mature. All the ESTs particularly existed in the 4(5) cell lines were sequenced then analysed by BLAST similarity search.ResultsWe gained 17 ESTs of 4(5)cells lines, three of them (gb|BE661968.1|,gb|BE661970.1|& gba|BE661982.1|)came from mouse cytochrome C oxidase CDS, Eps8mRNA,Hsp65mRNA respectively, others represented new genes. Conclusion DD-PCR was a feasible method for finding new expressed genes, mouse thymus might express more molecules than that we know now.