| 葡萄卷叶伴随病毒-3外壳蛋白的原核表达及其特异性抗血清的制备 |
| |
| 引用本文： | 徐章逸,王国平,刘亚萍,王彩霞,洪霓.葡萄卷叶伴随病毒-3外壳蛋白的原核表达及其特异性抗血清的制备[J].植物病理学报,2010,40(2):129-134. |
| |
| 作者姓名： | 徐章逸 王国平 刘亚萍 王彩霞 洪霓 |
| |
| 作者单位： | 1 湖北省作物病害监测和安全控制实验室, 武汉 430070;2 华中农业大学国家果树脱毒种质资源室内保存中心, 武汉 430070 |
| |
| 基金项目： | 科技部"863"课题资助项目 |
| |
| 摘 要： | 利用RT-PCR技术扩增得到了葡萄卷叶伴随病毒-3(Grapevine leafroll associated virus-3,GLRaV-3)中国分离物的外壳蛋白(coat protein,cp)基因。序列分析结果表明,cp基因的长度为942bp,与已报道的其它GLRaV-3分离物的cp核苷酸相似性为91%～99%,编码的氨基酸相似性为95%～100%。将此基因克隆到原核表达载体pET-28a(+)上,转化大肠杆菌BL21(DE3)plysS后用终浓度为1mmol/L的IPTG进行诱导表达,SDS-PAGE及Western blotting分析表明,cp在大肠杆菌中可表达出分子量约为35kDa的蛋白。纯化表达产物后免疫家兔制备抗血清。A蛋白酶联免疫吸附测定(PAS-ELISA)及斑点免疫结合测定(DBIA)结果显示,制备的特异抗血清可用于检测田间感病葡萄样品中的GLRaV-3。
|
| 关 键 词： | GLRaV-3 cp 基因克隆 原核表达 PAS-ELISA DBIA |
Prokaryotic expression of coat protein of Grapevine leafroll associated virus-3 and its special antiserum preparation |
| |
| XU Zhang-yi,WANG Guo-ping,LIU Ya-ping,WANG Cai-xia,HONG Ni.Prokaryotic expression of coat protein of Grapevine leafroll associated virus-3 and its special antiserum preparation[J].Acta Phytopathologica Sinica,2010,40(2):129-134. |
| |
| Authors: | XU Zhang-yi WANG Guo-ping LIU Ya-ping WANG Cai-xia HONG Ni |
| |
| Institution: | 1 Hubei Crop Disease Monitoring and Safety Control Key Laboratory, Wuhan 430070, China;2 National Indoor Conservation Center for Virus-free Gemplasms of Fruit Crops, Huazhong Agricultural University, Wuhan 430070, China |
| |
| Abstract: | The coat protein gene of a Chinese isolate of Grapevine leafroll associated virus-3 was amplified by RT-PCR. Sequence analysis showed that the cp was 942 bp,and it had different similarities with that of other reported GLRaV-3 isolates ranging 91%-99% at nucleotide sequence level and 95%-100% at amino acid sequence level. This gene was cloned into the prokaryotic expression vector pET-28a(+),then trans formed into E. coli BL-21 (DE3 plysS),and subsequently induced by IPTG at final concentration of 1 mmol/L.... |
| |
| Keywords: | GLRaV-3 cp PAS-ELISA DBIA |
| 本文献已被 CNKI 万方数据 等数据库收录！ |
| 点击此处可从《植物病理学报》浏览原始摘要信息 |
| 点击此处可从《植物病理学报》下载免费的PDF全文 |
|
