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| 人抵抗素基因全长的体外拼接及真核表达载体的构建 | |
| 引用本文： | 乔小芝,王宪锋,赵冬久,徐哲荣,俞玲娣,陈智,杨云梅. 人抵抗素基因全长的体外拼接及真核表达载体的构建[J]. 浙江大学学报(医学版), 2007, 36(6): 588-591,609 |
| 作者姓名： | 乔小芝 王宪锋 赵冬久 徐哲荣 俞玲娣 陈智 杨云梅 |
| 作者单位： | 1. 浙江大学医学院,附属第一医院干部病房,浙江,杭州,310003 2. 浙江大学医学院,第一附属医院传染病研究所,浙江,杭州,310003 3. 浙江大学医学院,解剖学教研室,浙江,杭州,310058 |
| 摘 要： | 目的:利用合成的寡核苷酸片段,在体外拼接人抵抗素(Resistin,Retn)基因的全长,并构建其真核表达载体。方法:根据已知抵抗素基因(GenBank:AF323081),设计并合成10条寡核苷酸片段,采用降落PCR方法进行拼接,获得预期大小的PCR产物后将其克隆入pSecTag2B载体进行测序确认。结果:降落PCR法扩增的特异条带与目的基因的大小一致。克隆载体测序结果与GenBank中已知序列完全符合。结论:降落PCR成功地拼接和扩增了人抵抗素全长基因,并构建了含有该基因的真核表达载体,为进一步研究该基因的功能奠定了实验基础。 |
| 关 键 词： | 激素类,异位 寡核苷酸类 聚合酶链反应 基因扩增 人Resistin基因 降落PCR 表达载体 |
| 文章编号： | 1008-9292（2007）06-0588-04 |
| 收稿时间： | 2007-05-22 |
| 修稿时间： | 2007-07-16 |
Assembly of full-length human resistin gene in vitro and construction of its eukaryotic expression vector |
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| QIAO Xiao-zhi, WANG Xian-feng, ZHAO Dong-jiu, et al. Assembly of full-length human resistin gene in vitro and construction of its eukaryotic expression vector[J]. Journal of Zhejiang University. Medical sciences, 2007, 36(6): 588-591,609 | |
| Authors: | QIAO Xiao-zhi WANG Xian-feng ZHAO Dong-jiu et al |
| Affiliation: | The First Affiliated Hospital, College of Medicine, Zhejiang University, Hangzhou 310003, China. qxz1971@hzcnc.com |
| Abstract: | Objective: To assemble the full-length of human resistin gene in vitro by using oligonucleotides and to construct its eukaryotic expression vector.Methods: According to the gene sequence of resistin(GenBank:AF323081),10 oligonucleotides were designed and synthesized,followed by a touch down PCR to assemble the full-length gene.The PCR products were cloned into pSecTag2B vector and confirmed by sequencing.Results: The band of PCR products and gene sequencing showed the insert fragment in pSecTag2B vector was identical to that as designed.Conclusion: The full-length of human resistin coding sequence was successfully assembled and amplified by touch down PCR,and a resistin-expressing eukaryotic vector was constructed. |
| Keywords: | Hormones ectopic Oligonucleotides Polymerase chain reaction Gene amplification Resistin gene Touch down PCR Expression vector |
| 本文献已被 CNKI 维普 万方数据 等数据库收录！ | |