| Fusion Expression and Purification of Marek’s disease virus Tegument Protein VP16 and Its Antibody Preparation |
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| 引用本文： | 邱亚峰,葛菲菲,冯秀丽,陈溥言.Fusion Expression and Purification of Marek’s disease virus Tegument Protein VP16 and Its Antibody Preparation[J].农业生物技术学报,2006,14(4):530-534. |
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| 作者姓名： | 邱亚峰 葛菲菲 冯秀丽 陈溥言 |
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| 作者单位： | 南京农业大学动物疫病诊断与免疫重点开放实验室,南京,210095 |
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| 基金项目： | 国家高技术研究发展计划(863计划) |
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| 摘 要： | 马立克氏病病毒(Marek’s disease virus ,MDV)UL48基因编码的蛋白与单纯疱疹病毒1型(Herpes simplex virus ,HSV)衣壳蛋白VP16为同源物，将其克隆入pET-32a载体中，获得pET-VP16, 转化Escherichia coli BL21感受态细胞，经IPTG诱导表达，SDS-PAGE电泳显示:融合蛋白获得可溶性表达，表达蛋白的相对分子量约为67 kD；将表达的融合蛋白过His·Bind柱，获得纯化的蛋白，将该蛋白免疫家兔，制备多克隆抗体。通过间接ELISA和Western blot鉴定制备的多克隆抗体的效价和特异性，结果表明，该抗体具有较高的特异性，间接ELISA效价大于2×10－5。
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| 关 键 词： | 马立克氏病病毒 融合表达 转录激活子 抗体 |
| 文章编号： | 1006-1304（2006）04-0530-03 |
| 收稿时间： | 2005-8-31 |
| 修稿时间： | 2006-3-30 |
Fusion Expression and Purification of Marek's disease virus Tegument Protein VP16 and Its Antibody Preparation |
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| QIU Ya-feng,GE Fei-fei,FENG Xiu-li,CHEN Pu-yan.Fusion Expression and Purification of Marek''''s disease virus Tegument Protein VP16 and Its Antibody Preparation[J].Journal of Agricultural Biotechnology,2006,14(4):530-534. |
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| Authors: | QIU Ya-feng GE Fei-fei FENG Xiu-li CHEN Pu-yan |
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| Abstract: | Gene UL48 of Marek’s disease virus(MDV), encoding the homologue of Herpes simplex virus (HSV) tegument protein VP16, was cloned into pET-32a to obtain pET-VP16. The recombinant vector pET-VP16 was transformed into Escherichia.coli BL21 and solubly expressed in LB medium as a fused protein of about 67 kD with induction of IPTG. And the fusion protein was purified by His·Bind chromatography column. Then immunization in rabbits with the fusion protein was used to obtain the high titer antibody against MDV VP16. Indirect ELISA indicated that the highest titer was over 2×10-5. Moreover, Western blotting analysis proved that the antibody was specific to VP16. |
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