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II型单纯疱疹病毒糖蛋白D的晶体结构解析
引用本文:陈竹君,逯光文,齐建勋,许翔,张娜,严景华,王荣富. II型单纯疱疹病毒糖蛋白D的晶体结构解析[J]. 生物工程学报, 2011, 27(10): 1499-1506
作者姓名:陈竹君  逯光文  齐建勋  许翔  张娜  严景华  王荣富
作者单位:1. 安徽农业大学生命科学学院,合肥 230036;中国科学院微生物研究所中国科学院病原微生物与免疫学重点实验室,北京 100101
2. 中国科学院微生物研究所 中国科学院病原微生物与免疫学重点实验室,北京,100101
3. 安徽农业大学生命科学学院,合肥,230036
基金项目:国家重点基础研究发展计划 (973计划) (No. 2010CB530104),安徽省教育厅自然科学基金重大项目 (No. KJ2010ZD04) 资助。
摘    要:II型单纯疱疹病毒 (HSV-2) 糖蛋白D (Glycoprotein D,gD) 在介导该病毒入侵到宿主细胞中起着关键作用。为了更好地研究gD在病毒侵入过程中的作用机制,利用杆状病毒表达系统表达了gD胞外部分区域 (1~285 aa),通过Ni-NTA亲和层析以及分子排阻层析纯化后,得到的带His标签的分泌型可溶蛋白,用悬滴法对该蛋白进行了晶体筛选,获得了高质量的晶体。晶体生长条件为0.1 mol/L Hepes缓冲液 (pH 7.2),5% (V/V) 2-甲基-2,4-戊二醇 (MPD),10%

关 键 词:杆状病毒表达系统,纯化,II型单纯疱疹病毒,糖蛋白D结晶
收稿时间:2011-03-05

X-ray diffraction analysis of glycoprotein D from Herpes simplex virus type 2
Zhujun Chen,Guangwen Lu,Jianxun Qi,Xiang Xu,Na Zhang,Jinghua Yan and Rongfu Wang. X-ray diffraction analysis of glycoprotein D from Herpes simplex virus type 2[J]. Chinese journal of biotechnology, 2011, 27(10): 1499-1506
Authors:Zhujun Chen  Guangwen Lu  Jianxun Qi  Xiang Xu  Na Zhang  Jinghua Yan  Rongfu Wang
Affiliation:College of Life Science, Anhui Agricultural University, Hefei 230036, China; CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China;CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China;CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China;College of Life Science, Anhui Agricultural University, Hefei 230036, China; CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China;CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China;CAS Key Laboratory of Pathogenic Microbiology and Immunology, Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China;College of Life Science, Anhui Agricultural University, Hefei 230036, China
Abstract:Glycoprotein D(gD) of Herpes simplex virus type 2(HSV-2) is a key factor mediating the entry of HSV-2 into host cells.In order to explain the mechanism underlying the gD-mediated receptor-binding and viral entry,we performed a structural study on HSV-2 gD.The ectodomain of the gD protein encompassing residues 1 to 285 was expressed by baculovirus-infected insect cells as a secreted soluble protein with a C-terminal hexa-his tag.The protein was then purified by affinity and size-exclusion chromatography.The ...
Keywords:expression by baculovirus-infected insect cells   purification   herpes simplex virus type 2   crystallization of glycoprotein D
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