| 重组大鼠谷氨酸脱羧酶载体的构建和功能分析(英文) |
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| 引用本文： | 刘建生,王倩,张继波,孔令菊,姚素艳,郑德宇,徐群渊. 重组大鼠谷氨酸脱羧酶载体的构建和功能分析(英文)[J]. 中国神经科学杂志, 2011, 0(6) |
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| 作者姓名： | 刘建生 王倩 张继波 孔令菊 姚素艳 郑德宇 徐群渊 |
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| 作者单位： | 辽宁医学院解剖学教研室;辽宁医学院附属第一医院放射科;辽宁医学院病理生理学教研室;首都医科大学神经科学研究所; |
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| 摘 要： | 目的谷氨酸脱羧酶2 (glutamic acid decarboxylase 2,GAD65) 是γ-氨基丁酸(gamma-aminobutyric acid, GABA)的合成酶。本研究拟构建重组大鼠GAD65基因的慢病毒载体(recombinant lentivirus-rGAD65,rLV-rGAD65),并在体内外分析其功能。方法用RT-PCR法克隆大鼠GAD65基因的cDNA 并亚克隆至慢病毒载体上,形成重组慢病毒质粒(rLV-GFP-rGAD65)。在包装质粒的帮助下,获得重组慢病毒颗粒(rLV-rGAD65)并检测其滴度。用rLV-rGAD65感染原代培养的大鼠肺成纤维细胞,并用免疫细胞化学和蛋白印迹法检测rGAD65在成纤维细胞中的表达,用高效液相法(high-performance liquid chromatograph, HPLC)检测培养上清中GABA的含量。在体内, rLV-rGAD65 定点注射到Sprague-Dawley大鼠的丘脑底核(subthalamic nucleus,STN)。用免疫组织化学和蛋白印迹法检测GAD65基因在STN中的表达水平,HPLC检测黑质网状部(su...
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| 关 键 词： | 大鼠谷氨酸脱羧酶2 慢病毒载体 基因克隆 帕金森病 |
Construction and functional activity of a recombinant vector expressing rat glutamic acid decarboxylase 65 |
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| Jian-Sheng Liu,Qian Wang,Ji-Bo Zhang,Ling-Ju Kong,Su-Yan Yao,De-Yu Zheng,Qun-Yuan Xu Departments of Anatomy , Pathophysiology,Liaoning Medical University,Jinzhou ,China. Construction and functional activity of a recombinant vector expressing rat glutamic acid decarboxylase 65[J]. Neuroscience Bulletin, 2011, 0(6) |
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| Authors: | Jian-Sheng Liu Qian Wang Ji-Bo Zhang Ling-Ju Kong Su-Yan Yao De-Yu Zheng Qun-Yuan Xu Departments of Anatomy Pathophysiology Liaoning Medical University Jinzhou China |
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| Affiliation: | Jian-Sheng Liu1,Qian Wang3,Ji-Bo Zhang1,Ling-Ju Kong1,Su-Yan Yao2,De-Yu Zheng1,Qun-Yuan Xu4 Departments of 1Anatomy and 2Pathophysiology,Liaoning Medical University,Jinzhou 121001,China 3Department of Radiology,First Affiliated Hospital of Liaoning Medical University,China 4Beijing Institute for Neuroscience,Capital Medical University,Beijing 100069,China |
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| Abstract: | Objective Glutamic acid decarboxylase 2(GAD65) is a gamma-aminobutyric acid(GABA) synthetase.This study aimed to construct a recombinant lentivirus-rGAD65(rLV-rGAD65) vector containing the cDNA of rat GAD65(rGAD65) and assess its functional activity in vitro and in vivo.Methods cDNA of rGAD65 was amplified by RT-PCR and subcloned into the LV vector,forming the rLV-GFP-rGAD65 plasmid.The recombinant lentivirus particles(rLVrGAD65) were packaged by the LV Helper-Free System and the titer was measured.Primary ... |
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| Keywords: | rat glutamic acid decarboxylase 2 lentivirus vector gene clone Parkinson's disease |
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