人MEPE/OF45基因的克隆及序列分析  

Cloning and Sequencing of Human MEPE/OF45 Gene

作　　者：张鸿声[1,2] 高宁[1] 绳纪坡[1] 于芳[1] 张宏达[1] 张勇[1,3] 胡宝成[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100850 [2]福建农林大学生物农药与化学生物学教育部重点实验室,福建福州350002 [3]福建农林大学农产品品质研究所,福建福州350002

出　　处：《生物技术通讯》2008年第6期787-790,共4页Letters in Biotechnology

基　　金：国家自然科学基金(30770651;30670616;30370441);国家重点基础研究发展计划(2005CB522506;2007CB914604);国家"十一五"科技支撑项目(2006BAI23B02)

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。Objective： To clone MEPE/OF45 full length gene in human ceils, 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 eDNA 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.

关 键 词：MEPE/OF45基因 克隆 序列分析 亚胺环己酮 HELA细胞 

分 类 号：Q78[生物学—分子生物学]