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以基因表达谱芯片对Ty2 1a免疫前后小鼠肠细胞 (包括肠粘膜上皮细胞和肠上皮间淋巴细胞 )基因表达的差异性进行研究比较。将 490条经抑制消减杂交法筛选出的与小鼠Ty2 1a免疫相关的cDNA制备成表达谱芯片 ;利用免疫前后小鼠肠细胞的mRNA通过逆转录方法 ,将Cy3和Cy5两种荧光分别标记到两种组织的cDNA上 ,制备成cDNA探针 ,并与表达谱芯片进行杂交及扫描 ,单点重复 2次实验 ,通过计算机数据处理判定基因是否在上述两种细胞群中有表达差异。筛选出差异表达基因共 98条 ,其中 92条为表达上调基因 ,6条为表达降低基因。提示 ,基因表达谱芯片技术是高通量进行基因表达模式研究的方法 ,可同时定量研究大量的基因表达水平 ,从而鉴定可能参与免疫的基因。
Differences in gene expression profiles of mouse intestinal cells (including intestinal mucosal epithelial cells and intestinal interstitial lymphocytes) before and after Ty2 1a immunization were compared using gene expression microarray. Four hundred and ninety-two cDNAs that were screened by inhibition subtractive hybridization were immunized with mouse Ty2 1a to prepare expression profiling chips. Two kinds of fluorescence of Cy3 and Cy5 were labeled by mRNA of mouse intestinal cells before and after immunization by reverse transcription CDNA of both tissues were prepared and cDNA probes were prepared. The cDNA probes were hybridized and scanned with the expression microarray. Single-point repeat experiments were performed twice to determine whether the genes were expressed in the two cell populations by computer data processing. A total of 98 differentially expressed genes were screened, of which 92 were up-regulated genes and 6 were down-regulated genes. Prompted, gene expression microarray technology is a high-throughput method of gene expression profiling that can simultaneously quantify a large number of gene expression levels to identify genes that may be involved in immunity.