【摘要】 目的:克隆人MEPE/OF45全长基因,为进一步研究MEPE/OF45在DNA损伤应答中的作用及特异的信号通路奠定基础。方法:选取人宫颈癌细胞HeLa为靶细胞,从中提取总RNA,设计扩增人MEPE/OF45基因片段的特异引物,进行RT-PCR分析;用蛋白质合成抑制剂亚胺环己酮(CHX)对细胞进行处理,观察处理前后MEPE/OF45基因的表达情况。结果:在经CHX处理的HeLa细胞中扩增到了MEPE/OF45基因片段,随后利用重叠PCR方法获得了人MEPE/OF45全长基因,并经序列分析确证。结论:获得了人MEPE/OF45基因片段及全长基因,为阐明MEPE/OF45在细胞中复杂的功能提供了线索。更多还原

【Abstract】 Objective: To clone MEPE / OF45 full length gene in human cells, so as to lay a foundation for studying the role of MEPE / OF45 in DNA damage responds and the specific signal pathways. Methods: Total RNA was extracted from human HeLa cells, the primers to amplify human MEPE / OF45 gene were designed, and cDNA was amplified by RT-PCR. The expressions of MEPE / OF45 gene fragments were observed in the cells after cycloheximide(CHX) treatment to inhibite protein synthesis. Results: Human MEPE / OF45 gene fragments were obtained after CHX treatment, full length gene was amplified by PCR, and this gene is homologous sequence with human MEPE / OF45 gene by sequencing analysis. Conclusions: Human MEPE/OF45 gene fragments and full length gene were obtained, it provides a clue to elucidate MEPE/ OF45 complicated function in the cells.更多还原