| 基于人脂肪组织获取基因构建pET28a-leptin(瘦素)重组子 |
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| 引用本文： | 文辉才,柳大烈,巫国辉,单磊. 基于人脂肪组织获取基因构建pET28a-leptin(瘦素)重组子[J]. 中国神经再生研究, 2009, 13(7): 1317-1319 |
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| 作者姓名： | 文辉才 柳大烈 巫国辉 单磊 |
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| 作者单位： | 南方医科大学珠江医院,南方医科大学珠江医院整形外科,江西省人民医院整形颌面外科,南方医科大学珠江医院整形外科 |
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| 基金项目： | 江西省卫生厅重大招标计划(2003 07)资助* |
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| 摘 要： | 目的：构建pET28a-leptin重组子。方法：取人脂肪组织，RT-PCR得到瘦素基因后通过DNA重组技术克隆至pMD18T载体与pET28a原核表达载体，EcoRⅠ+XhoⅠ双酶切及测序鉴定。结果：扩增得到520bp目的基因；所构建重组瘦素原核表达载体pET28a-leptin经双酶切电泳见520bp目的条带，DNA测序与相应基因序列同源性达100%。结论：成功构建pET28a-leptin重组子，为后续研究提供了基础和物质保障。
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| 关 键 词： | 瘦素；重组；脂肪 |
| 收稿时间： | 2009-02-03 |
| 修稿时间： | 2009-02-03 |
Construction of recombinant pET28a-leptin based on human adipose tissues |
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| Abstract: | Objective: To construct recombinant pET28a-leptin. Methods: Human adipose tissue was used for amplifying leptin gene by RT-PCR. The gene was cloned into pMD18T and pET28a to construct recombinant which was identified by double enzyme digest and DNA sequencing. Results: The gene amplified was 520bp. Meanwhile, the 520bp fragment was got by double enzyme digest, between which and human obesity gene the sequence homology is a hundred per cent. Conclusion: The recombinant pET28a-leptin was constructed, which provide material for further study |
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