| 靶向性DNA甲基化酶在pET32a原核表达载体中的表达和鉴定 |
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| 引用本文： | 吴琳,邢克飞,张治国,张健,刘新平,药立波,李福洋. 靶向性DNA甲基化酶在pET32a原核表达载体中的表达和鉴定[J]. 现代生物医学进展, 2008, 8(9): 1659-1661,1630 |
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| 作者姓名： | 吴琳 邢克飞 张治国 张健 刘新平 药立波 李福洋 |
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| 作者单位： | 1. 第四军医大学基础部生物化学与分子生物学教研室,陕西西安,710032
2. 第四军医大学预防医学系流行病学教研室,陕西西安,710032 |
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| 摘 要： | 目的:在pET32a原核表达载体中表达融合myc-6his标签的靶向性甲基化酶B1-3a并进行鉴定。方法:以含有B1-3a基因的pcDNA4.0-B1-3a-myc-6his质粒为模板,通过PCR的方法扩增获得融合有myc-6his标签序列的目的区段B1-3a,然后克隆入表达载体pET32a;以SDS-PAGE和Western blot方法对表达产物进行鉴定。结果:表达产物中在分子量43kD左右可见与目的蛋白分子量相符的条带,该条带可被6his标签单克隆抗体特异识别。结论:正确构建了靶向性甲基化酶B1-3a的原核表达载体,靶向性甲基化酶能够在pET32a中成功表达。
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| 关 键 词： | DNA甲基化酶 基因 靶向性 原核表达载体pET32a |
Expression and Identification of Targeting Methyltransferase in pET32a Expression Vector |
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| WU Lin,XING Ke-fei,ZHANG Zhi-guo,ZHANG Jian,LIU Xin-Ping,YAO Li-bo,LI Fu-yang. Expression and Identification of Targeting Methyltransferase in pET32a Expression Vector[J]. Progress in Modern Biomedicine, 2008, 8(9): 1659-1661,1630 |
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| Authors: | WU Lin XING Ke-fei ZHANG Zhi-guo ZHANG Jian LIU Xin-Ping YAO Li-bo LI Fu-yang |
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| Abstract: | Objective:To clone and express the targeting methyltranferase B1-3a gene fused with myc-6his tag in prokaryotic ex-pression system and to identify the protein of fused expression.Methods:The B1-3a gene was amplified from pcDNA4.0-B1-3a-his-6myc plasmid with PCR method and cloned into the expression vector pET32a.The expression product was analyzed with SDS-PAGE and Western blot.Results:A positive protein band with molecular weight about 43 kD was found in the expression product, which is accordant with interest protein, and this band could be specifically recognized by anti-6his monoclonal antibody.Conclusion:The targeting DNA methyltransferase B1-3a was cloned into the expression vector correctly and expressed successfully in prokaryotic ex-pression system. |
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| Keywords: | DNA methyltransferase gene targeting prokaryotic expression vector pET32a |
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