N-甲基-N′-硝基-N-亚硝基胍引起vero细胞基因表达改变及有关cDNA片段的初步鉴定

运用mRNA差异显示(DD-PCR)技术，通过26 对锚定及任意引物组合显示基因的差异表达情况，分离了7 个表达有差异的片段.其中3 个片段的相关基因属于对N-甲基-N′-硝基-N-亚硝基胍(MNNG)处理的初级反应基因，2个属于次级反应基因.另有2 个片段的差异表达仅在蛋白合成抑制剂环己亚胺（CHM）合并MNNG处理时才出现.反向缝隙印迹杂交分析印证了2 个片段在DD-PCR中发现的改变，同时分析的本实验室分离的25个片段中，8 个片段的变化被验证与DD-PCR中的改变一致.序列分析结果显示这些片段与许多基因有高同源性，其中包括一些参与信息传导的基因.

Differential gene expression induced by N-methyl-N′-nitro-N-nitrosoguanidine in vero cell and tentative identification of relevant cDNA fragments

Differential gene expression was examined by means of DD- PCR with 26 pairs of anchored and arbitrary primers. Seven differentially expressed fragments were isolated after N-methyl-N′-nitro-N-nitrosoguanidine(MNNG) exposure, among them, three were classified as primary response genes, two as secondary response genes, and another two differentially expressed fragments were observed only after combined protein synthesis inhibitor(cycloheximide, CHM) and MNNG exposure. The expression patterns of two fragments were confirmed by reverse slot blot analysis as well as other eight of twenty-five fragments isolated simultaneously in this lab. Sequencing data showed that these fragments had high homology with several genes, including some genes participating in signal transduction.