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病毒感染植物汁液的电印迹放射免疫测定法——一种检测植物病毒的强大新技术。

Electro-blot radioimmunoassay of virus-infected plant sap - a powerful new technique for detecting plant viruses.

作者信息

O'Donnell I J, Shukla D D, Gough K H

出版信息

J Virol Methods. 1982 Feb;4(1):19-26. doi: 10.1016/0166-0934(82)90050-7.

Abstract

A new technique for detecting viruses in plant sap is described. It consists of sodium dodecyl sulphate-polyacrylamide gel electrophoresis of the infected plant sap, electrophoretic transfer of protein bands to activated paper by the Electro-Blot technique, the subsequent probing of the viral coat protein band by specific antiserum (prepared against intact virus), and detection of immune complex with 125 I-labelled protein A. The technique successfully detected tobacco mosaic virus at a sap dilution of 1 : 10,000, four strains of sugarcane mosaic virus (a potyvirus) in their perennial hosts infected for about 4 years, and five different isolated of potato leaf roll virus (a luteovirus). The latter virus occurs in extremely low concentration and is difficult to detect by the other known methods.

摘要

本文描述了一种检测植物汁液中病毒的新技术。该技术包括对受感染植物汁液进行十二烷基硫酸钠-聚丙烯酰胺凝胶电泳,通过电印迹技术将蛋白带电泳转移至活化纸上,随后用特异性抗血清(针对完整病毒制备)探测病毒衣壳蛋白带,并用¹²⁵I标记的蛋白A检测免疫复合物。该技术成功检测到了稀释度为1:10000的烟草花叶病毒、在多年生寄主中感染约4年的4种甘蔗花叶病毒株系(一种马铃薯Y病毒属病毒)以及5种不同分离株的马铃薯卷叶病毒(一种黄症病毒属病毒)。后一种病毒浓度极低,用其他已知方法难以检测。

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