| 人类p27kip1基因正、反义真核表达质粒的构建 |
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| 引用本文： | 张冬梅,程纯,严美娟,陆建新.人类p27kip1基因正、反义真核表达质粒的构建[J].南通医学院学报,2005,25(1):4-6. |
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| 作者姓名： | 张冬梅 程纯 严美娟 陆建新 |
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| 作者单位： | 南通大学微生物与免疫学教研室,南通,226001 |
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| 基金项目： | 江苏省高校自然科学研究项目(04KJB320114),江苏省2004年度研究生创新计划项目(xm04-66) |
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| 摘 要： | 目的：克隆人类p27^kip1基因，构建其正、反义真核表达质粒。方法：采用逆转录聚合酶链式反应(RT—PCR)技术，从人类伯基特淋巴瘤细胞株Raji中扩增出人类p27^kip1 cDNA全长序列，通过DNA重组技术将该基因重组于pcDNA3．1真核表达载体上，构建p27^kip1正、反义表达质粒。通过酶切电泳分析及DNA测序的方法对重组表达质粒进行鉴定。结果：通过酶切鉴定及测序分析证实，本实验构建的重组表达质粒目的基因片段为人类p27^kip1 cDNA。结论：本实验成功地克隆了人类p27kipl基因并构建了其正、反义真核表达质粒，为进一步研究p27^kip1在淋巴瘤发生中的意义奠定了基础。
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| 关 键 词： | p27^kip1 基因克隆 基因重组 非霍奇金淋巴瘤 逆转录聚合酶链反应 |
| 文章编号： | 1000-2057(2005)01-0004-03 |
| 修稿时间： | 2004年10月25 |
Construction of p27kip1 gene sense and antisense eukaryotic expressive plasmids |
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| ZHANG Dongmei,CHENG Chun,YAN Meijuan,et al.Construction of p27kip1 gene sense and antisense eukaryotic expressive plasmids[J].ACTA Academiae Medicinae Nantong,2005,25(1):4-6. |
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| Authors: | ZHANG Dongmei CHENG Chun YAN Meijuan |
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| Abstract: | Objective:To clone p27 kip1 gene and to construct its sense and antisense expressive plasmids.Methods:Human p27 kip1 complete cDNA was amplified by RT-PCR method from human Burkitt lymphoma cell line Raji , and was reconstructed into the eukaryotic expressive vector pcDNA3.1 to form the p27 kip1 sense or antisense expressive plasmid. The reconstructed DNA was identified by enzyme digestion and DNA sequencing.Results:The cloned DNA was confirmed to be human p27 kip1 cDNA.Conclusion:In this study we successfully cloned p27 kip1 gene and constructed its sense and antisense expressive plasmids, which provided the foundation for further studying the significance of p27 kip1 in lymphoma. |
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