| 小鼠Prx2正义和反义真核表达载体的构建与表达 |
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| 引用本文： | 杨书红,吕兴,罗爱月,赖志文,丁婷,韩凌斐,卢运萍,王世宣.小鼠Prx2正义和反义真核表达载体的构建与表达[J].现代生物医学进展,2011,11(16):3005-3008. |
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| 作者姓名： | 杨书红 吕兴 罗爱月 赖志文 丁婷 韩凌斐 卢运萍 王世宣 |
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| 作者单位： | 1. 华中科技大学附属同济医院妇产科 湖北武汉 430030
2. 郧阳医学院附属太和医院肝胆外科 湖北十堰 442000 |
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| 基金项目： | 卫生部公益性行业科研专项项目，教育部高等学校博士点科研基金，国家自然科学基金 |
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| 摘 要： | 目的:构建小鼠Prx2正义及反义真核表达载体pEGFP-Nl-sPrx2、pEGFP-N1-aPrx2并在小鼠卵巢颗粒细胞中表达。方法:应用RT-PCR技术,从小鼠卵巢颗粒细胞提取的总RNA中,获得Prx2基因编码序列的正义及反义片段,克隆至真核表达载体pEGFP-N1中,对重组质粒进行酶切和测序鉴定后,以脂质体介导法转染至体外培养的未成熟小鼠颗粒细胞,通过荧光显微镜观察和Westernblot检测其在颗粒细胞中的表达改变。结果:酶切和测序证明重组真核表达载体pEGFP-N1-sPrx2、pEGFP-Nl-aPrx2构建成功,荧光显微镜观察及Western blot确认目标蛋白在颗粒细胞中表达增强或减弱。结论:成功构建小鼠Prx2基因正义及反义真核表达载体pEGFP-N1-sPrx2、pEGFP-N1-aPrx2并在小鼠卵巢颗粒细胞中表达。
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| 关 键 词： | Prx2 正义/反义 真核表达载体 颗粒细胞 小鼠 |
Construction and Expression of Mouse Prx2 Sense or Antisense Eukaryotic Expression Vectors |
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| YANG Shu-hong,LV Xing,LUO Ai-yue,LAI Zhi-wen,DING Ting,HAN Ling-fei,LU Yun-ping,WANG Shi-xuan.Construction and Expression of Mouse Prx2 Sense or Antisense Eukaryotic Expression Vectors[J].Progress in Modern Biomedicine,2011,11(16):3005-3008. |
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| Authors: | YANG Shu-hong LV Xing LUO Ai-yue LAI Zhi-wen DING Ting HAN Ling-fei LU Yun-ping WANG Shi-xuan |
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| Institution: | 1 Department of Obstetrics and Gynecology,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan,Hubei 430030,PR China;2 Hepatobiliary Department of Taihe Hospital,Yunyang Medical College,Shiyan,Hubei 442000,China) |
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| Abstract: | Objective:To construct the sense or antisense mouse Prx2 eukaryotic expression vectors pEGFP-N1-sPrx2 and pEGFP-N1-aPrx2.Methods:Sense and antisense Prx2 full length gene were amplified by RT-PCR from the total RNA of the granulosa cells(GCs) of the mouse ovary,then they were cloned into eukaryotic expression vector pEGFP-N1.After the comformation with PCR, restriction endonuclease digestion and the nucleotide sequencing,the recombinant vectors were transfected into the the GCs which were isolated from the ... |
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| Keywords: | Prx2 Sense/Antisense Eukaryotic expression vector Granulosa cells Mouse |
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