| 人类博卡病毒(HBoV)非结构蛋白NS1基因的克隆、原核表达及多克隆抗体的制备 |
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| 引用本文： | 孙彬,李京京,欧阳锦凤,陈莹,李毅.人类博卡病毒(HBoV)非结构蛋白NS1基因的克隆、原核表达及多克隆抗体的制备[J].现代生物医学进展,2010,10(6). |
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| 作者姓名： | 孙彬 李京京 欧阳锦凤 陈莹 李毅 |
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| 作者单位： | 华中师范大学生命科学学院,湖北,武汉,430079 |
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| 基金项目： | 国家自然科学基金(Nos.30670081) |
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| 摘 要： | 目的:克隆、表达、纯化人类博卡病毒(HBoV)非结构蛋白NS1,制备抗NS1多克隆抗体。方法:利用PCR扩增HBoV非结构蛋白NS1基因,将其克隆至pMAL-c2X表达载体上,重组质粒转化大肠杆菌DH10B,IPTG诱导表达。表达的融合蛋白经Amylose Resin亲和层析柱纯化后,免疫新西兰大白兔制备多克隆抗体。用间接ELISA法检测抗体效价。结果:原核表达融合蛋白MBP-NS1,并获得了其多克隆抗体,抗体效价达到1∶32000。结论:在原核表达系统中表达、纯化了融合蛋白,制备抗NS1多克隆抗体,为进一步研究该病毒非结构蛋白基因的转录和翻译机制提供可靠的工具。
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| 关 键 词： | 人类博卡病毒(HBoV) 非结构蛋白NS1 克隆 原核表达 多克隆抗体 |
Cloning of Non-structural Protein NS 1 Gene from Human Bocavirus and Preparation of Its Polyclonal Antibody |
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| SUN Bin,LI Jing-jing,OUYANG Jin-feng,CHEN Ying,LI Yi.Cloning of Non-structural Protein NS 1 Gene from Human Bocavirus and Preparation of Its Polyclonal Antibody[J].Progress in Modern Biomedicine,2010,10(6). |
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| Authors: | SUN Bin LI Jing-jing OUYANG Jin-feng CHEN Ying LI Yi |
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| Institution: | SUN Bin,LI Jing-jing,OUYANG Jin-feng,CHEN Ying,LI Yi (College of Life Sciences,huazhong normal university,wuhan,hubei 430079,China) |
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| Abstract: | Objective: To make polyclonal antibody by cloning and expressing the nonstructural protein gene NS1 of the Human bocavirus (HBoV) in Escherichia coli DH10B.Methods: NS1 gene of the HBoV was amplified by PCR and inserted into prokaryotic expression vector pMAL-c2X which contained the maltose binding protein (MBP),and then it was expressed in Escherichia coli DH10B strain.The target fusion protein was purified by Amylose affinity chromatography,and was used to immunize the New Zealand white rabbit for prepari... |
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| Keywords: | Human bocavirus nonstructural protein cloning prokaryotic expression polyclonal antibody |
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