| 李属坏死环斑病毒CP基因在大肠杆菌中的表达及其抗血清制备 |
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| 引用本文： | 陈招荣,马云霞,范在丰,李怀方.李属坏死环斑病毒CP基因在大肠杆菌中的表达及其抗血清制备[J].植物检疫,2006,20(2):73-75. |
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| 作者姓名： | 陈招荣 马云霞 范在丰 李怀方 |
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| 作者单位： | 中国农业大学植物病理系植物病毒实验室,北京,100094 |
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| 基金项目： | 引进国际先进农业科技计划(948计划) |
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| 摘 要： | 本实验用RT—PCR的方法获得李属坏死环斑病毒的印基因，并将其连接到表达载体pET-22b（＋）上，得到重组质粒pET—PNRSVCP，转化大肠杆菌BL21（DE3）后，用IPTG进行诱导表达。SDSPAGE和Westernblot分析结果表明，印基因在大肠杆菌中获得了高效表达，获得的融合蛋白分子量约为29．0kDa。将该融合蛋白免疫兔子，获得PNRSV特异性抗血清。酶联法（ELISA）测定的效价为1／1024。为用血清学方法检测该病毒提供了基础条件。
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| 关 键 词： | 李属坏死环斑病毒 外壳蛋白 原核表达 抗血清 |
| 收稿时间： | 2005-08-29 |
| 修稿时间： | 2005-08-29 |
Prokaryotic expression of the cp gene and antiserum preparation of Prunus necrotic ringspot virus. |
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| Chen Zhaorong,Ma Yunxia,Fan Zaifeng,Li Huaifang.Prokaryotic expression of the cp gene and antiserum preparation of Prunus necrotic ringspot virus.[J].Plant Quarantine,2006,20(2):73-75. |
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| Authors: | Chen Zhaorong Ma Yunxia Fan Zaifeng Li Huaifang |
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| Institution: | Phytovirology Lab. Plant Pathology Dept. China Agricultural University, Beijing 100094, China |
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| Abstract: | The coat protein (CP) gene of Prunus necrotic ringspot virus (PNRSV) was amplified by RT-PCR, and ligated to the expression vector pET-22b(+). The recombinant plasmid pET-WVMVCP was transformed into E. coli BL21(DE3) and then induced by IPTG. It was showed that the CP gene was highly expressed by SDS-PAGE and Western blot analysis. The molecular weight of the recombinant protein was about 29.0 kDa. Antiserum with high specificity was produced after the rabbit was immunized with purified recombinant protein, and the titer was determined to be 1/1024 by antigen coating plate- ELISA(ACP-ELISA). It provides the basic condition for serological detection of PNRSV. |
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| Keywords: | ELISA |
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