| 小鼠HSP27基因腺病毒表达载体的构建与鉴定 |
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| 引用本文： | 刘金娟,马 翔,刁飞扬,李 梅,薛 凯,崔毓桂,刘嘉茵.小鼠HSP27基因腺病毒表达载体的构建与鉴定[J].南京医科大学学报,2009,29(7):944-948. |
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| 作者姓名： | 刘金娟 马 翔 刁飞扬 李 梅 薛 凯 崔毓桂 刘嘉茵 |
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| 作者单位： | 刘金娟,LIU Jin-juan(中国药科大学生命科学与技术学院,江苏,南京,210038;南京医科大学第一附属医院生殖医学中心,江苏南京210029);马翔,刁飞扬,李梅,薛凯,崔毓桂,刘嘉茵,MA Xiang,DIAO Fei-yang,LI Mei,XUE Kai,CUI Yu-gui,LIU Jia-yin(南京医科大学第一附属医院生殖医学中心,江苏南京,210029)
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| 基金项目： | 国家自然科学基金，江苏省科教兴卫工程基金 |
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| 摘 要： | 目的:为深入研究热休克蛋白27(heat shock protein 27,HSP27)的生理与病理功能,应用PAdEasy腺病毒载体系统构建带荧光蛋白的小鼠HSP27重组腺病毒,并进行鉴定.方法:采用基因技术,将目的基因克隆至穿梭质粒pAdTrack-CMV中,通过PCR、序列测定、酶切鉴定;利用PAdEasy系统进行重组,在293细胞中包装和扩增,然后体外感染小鼠精原母细胞,Westernblot检测HSP27蛋白表达.结果:测序和酶切鉴定重组腺病毒质粒构建正确;经293细胞包装后可观察到绿色荧光,收获高滴度重组腺病毒体外感染小鼠精原母细胞,并且可正确表达HSP27.结论:应用PAdEasy系统重组技术成功构建了小鼠HSP27带荧光蛋白腺病毒载体,并在小鼠精母细胞系进行体外有效表达,为进一步研究HSP27的功能奠定基础.
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| 关 键 词： | 腺病毒 同源重组 |
| 收稿时间： | 2/4/2009 12:00:00 AM |
Construction and identification of recombinant adenovirus vector carrying mouse HSP27 |
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| LIU Jin-juan,MA Xiang,DIAO Fei-yang,LI Mei,XUE Kai,CUI Yu-gui and LIU Jia-yin.Construction and identification of recombinant adenovirus vector carrying mouse HSP27[J].Acta Universitatis Medicinalis Nanjing,2009,29(7):944-948. |
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| Authors: | LIU Jin-juan MA Xiang DIAO Fei-yang LI Mei XUE Kai CUI Yu-gui and LIU Jia-yin |
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| Institution: | 1Department of Life Science and Technology;China Pharmaceutical University;Nanjing 210038;2Center of Clinical Reproductive Medicine;the First Affiliated Hospital of NJMU;Nanjing 210029;China |
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| Abstract: | Objective:To investigate the physiological and pathological function of HSP27 by construct and identify the recombinant adenoviruses containing green fluorescence protein(GFP)and mouse HSP27 gene with PAdEasy system. Methods:The coding region of HSP27 was subcloned into the shuttle vector pAdTrack-CMV to form pAdTrack-CMV-HSP27. The identification was performed by PCR,sequencing and restriction digest. Chemical transformation of the plasmid pAdEasy-1 into E. coli BJ5183 strain was performed to prepare BJ518... |
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| Keywords: | HSP27 PCOS |
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