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| 人MEPE/OF45基因的克隆及序列分析 | |
| 引用本文： | 张鸿声,高宁,绳纪坡,于芳,张宏达,张勇,胡宝成. 人MEPE/OF45基因的克隆及序列分析[J]. 生物技术通讯, 2008, 19(6): 787-790 |
| 作者姓名： | 张鸿声 高宁 绳纪坡 于芳 张宏达 张勇 胡宝成 |
| 作者单位： | 1. 军事医学科学院生物工程研究所,北京,100850;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 2. 军事医学科学院生物工程研究所,北京,100850 3. 军事医学科学院生物工程研究所,北京,100850;福建农林大学,农产品品质研究所,福建,福州,350002 |
| 基金项目： | 国家自然科学基金，国家重点基础研究发展计划(973计划)，国家科技支撑项目 |
| 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。 |
| 关 键 词： | MEPE/OF45基因 克隆 序列分析 亚胺环己酮 HeLa细胞 |
Cloning and Sequencing of Human MEPE/OF45 Gene |
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| ZHANG Hong-Sheng,GAO Ning,SHENG Ji-Po,YU Fang,ZHANG Hong-Da,ZHANG Yong,HU Bao-Cheng. Cloning and Sequencing of Human MEPE/OF45 Gene[J]. Letters in Biotechnology, 2008, 19(6): 787-790 | |
| Authors: | ZHANG Hong-Sheng GAO Ning SHENG Ji-Po YU Fang ZHANG Hong-Da ZHANG Yong HU Bao-Cheng |
| Affiliation: | ZHANG Hong-Shen, GAO Ning, SHENG Ji-Po, YU Fang,ZHANG Hong-Da,ZHANG Yong,HU Bao-Cheng( 1. Beijing Institute of Biotechnology, Bcijing 100850; 2. a. Key Laboratory of Biopesticide and Chemical Biology; b. Agricultural Product Quality Institute; Fujian Agriculture and Forestry University, Fuzhou 350002; China) |
| Abstract: | 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. |
| Keywords: | MEPE/OF45 gene cloning sequencing analysis cycloheximide HeLa cells |
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