| pcDNA3.1(+)-VEGF165载体的构建及表达蛋白特性分析 |
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| 引用本文： | 邱明,肖明朝,苟欣,杨钰兴.pcDNA3.1(+)-VEGF165载体的构建及表达蛋白特性分析[J].生物技术通报,2010(2). |
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| 作者姓名： | 邱明 肖明朝 苟欣 杨钰兴 |
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| 作者单位： | 重庆医科大学附属第一医院泌尿外科,重庆,400016 |
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| 基金项目： | 重庆市自然科学基金计划项目 |
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| 摘 要： | 构建含人VEGF165基因的重组真核表达质粒,并对其表达蛋白特性进行初步分析。将合成的VEGF165基因序列克隆至pCR2.1-TOPO载体,测序鉴定证实基因碱基序列无误后,再克隆至真核表达载体pcDNA3.1(+),构建成pcD-NA3.1(+)-VEGF165重组质粒。利用DNAstar软件分析基因序列并翻译成氨基酸序列,再用ExPASy protscale和Protean软件分析其疏水特性、蛋白二级结构。经基因测序、酶切鉴定和PCR鉴定证实pcDNA3.1(+)-VEGF165重组质粒构建成功。Ex-PASy protscale和Protean软件分析表明VEGF165蛋白具有较好的水溶性。本研究为VEGF165的基因治疗应用和VEGF165蛋白直接治疗勃起功能障碍奠定了基础。
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| 关 键 词： | 血管内皮生长因子 基因克隆 序列测定 基因表达 |
Construction of pcDNA3.1(+)-VEGF165 and Analysis of the Expressed Protein |
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| Qiu Ming,Xiao Mingzhao,Gou Xin,Yang Yuxing.Construction of pcDNA3.1(+)-VEGF165 and Analysis of the Expressed Protein[J].Biotechnology Bulletin,2010(2). |
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| Authors: | Qiu Ming Xiao Mingzhao Gou Xin Yang Yuxing |
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| Institution: | Qiu Ming Xiao Mingzhao Gou Xin Yang Yuxing(Department of Urology,the First Affiliated Hospital,Chongqing Medical University,Chongqing 400016) |
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| Abstract: | It was to construct the pcDNA3.1(+)-VEGF165 expression vector and analyze the expressed protein.VEGF165 gene was cloned to pCR2.1-TOPO vector and then cloned into the vector pcDNA3.1(+)after sequencing.Two softwares ExPASy protscale and Protean were used to analyze the hydrophobic character and secondary structure of VEGF165 protein.Results indicated that pcDNA3.1(+)-VEGF165 was successfully constructed and confirmed by gene sequencing and PCR identification.Analysis with ExPASy protscale and Protean showed... |
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| Keywords: | Vascular endothelial growth factor Gene cloning Gene sequencing Gene expression |
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