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针对马铃薯Y病毒重组外壳蛋白的多克隆抗体的制备。

Production of polyclonal antibodies to a recombinant coat protein of potato virus Y.

作者信息

Folwarczna J, Plchová H, Moravec T, Hoffmeisterová H, Dedic P, Cerovská N

机构信息

Intitute of Experimental Botany, Academy of Sciences of the Czech Republic, 160 00, Prague, Czech Republic.

出版信息

Folia Microbiol (Praha). 2008;53(5):438-42. doi: 10.1007/s12223-008-0067-1. Epub 2008 Dec 16.

Abstract

The gene encoding the coat protein (CP) of a potato virus Y (PVY) was cloned into expression vector pMPM-A4Omega. PVY CP was expressed in Escherichia coli and the purified recombinant protein was used for raising rabbit polyclonal antibodies. The sera and antibodies were tested for the detection of PVY in the laboratory host Nicotiana tabacum cv. Petit Havana SR1 and in various cultivars of the natural host Solanum tuberosum by ELISA as well as by Western blots. The antibodies can be used for the detection of the whole strain spectrum of PVY by indirect plate trapped antigen ELISA and Western blot, but not by double antigen sandwich ELISA.

摘要

将马铃薯Y病毒(PVY)外壳蛋白(CP)的编码基因克隆到表达载体pMPM - A4Omega中。PVY CP在大肠杆菌中表达,纯化后的重组蛋白用于制备兔多克隆抗体。通过酶联免疫吸附测定(ELISA)以及蛋白质免疫印迹法,对血清和抗体在实验室寄主烟草品种 Petit Havana SR1以及天然寄主马铃薯的多个品种中检测PVY的能力进行了测试。这些抗体可通过间接板包被抗原ELISA和蛋白质免疫印迹法用于检测PVY的全株系谱,但不能通过双抗原夹心ELISA法检测。

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