伤寒减毒活疫苗Ty21a免疫前后小鼠肠细胞差异表达的基因

Study on the Gene Expressions in Murine Intestinal Cells before and after Immunization with Attenuated Living Vaccine of Salmonella typhi Ty21a by cDNA Microarray

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。

The expressions of genes in murine intestinal cells (including epithelial cells and intraepithelial lymphocytes) before and after immunization with Ty21a vaccine were studied by using the cDNA microarray. DNA microarrays were made by spotting PCR products of 490 Ty21a-associated cDNA of mice selected by SSH into a specially treated glass slide. The probe used was prepared by labeling intestinal mRNA of mice before and after immunization with Cy-3dUTP and Cy5-dUTP by means of reverse transcription. The arrays were then hybridized with cDNA probe,and the fluorescent signals scanned.Data thus obtained were analyzed from two repeated experiments.All together,98 differenially expressed genes were scanned out after immunization,in which 92 were up-regulated genes and 6 were down-regulated genes. It suggests that high throughput cDNA microarray is a credible method to profile changes in gene expression,and can be used for the quantitative studies of large scale of gene expression.