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在昆虫和植物宿主中对水稻矮缩植物呼肠孤病毒蛋白进行免疫检测。

Immunodetection of rice dwarf phytoreoviral proteins in both insect and plant hosts.

作者信息

Suzuki N, Sugawara M, Kusano T, Mori H, Matsuura Y

机构信息

Laboratory of Plant Genetic Engineering, Akita Prefectural College of Agriculture, Japan.

出版信息

Virology. 1994 Jul;202(1):41-8. doi: 10.1006/viro.1994.1320.

Abstract

Peptides encoded by truncated (S1 and S2) or full-length (S3 to S11) cDNAs of 11 of the 12 rice dwarf phytoreovirus (RDV) genome segments were expressed in a baculovirus vector system. Antibodies raised against each of the expressed peptides were used as probes to detect the authentic proteins encoded by the RDV open reading frame. Polypeptides identified as gene products of S1 to S11 in both RDV-infected rice leaf and leafhopper (Nephotettix cincticeps) homogenates were the P1 minor core (170 kDa), P2 (130 kDa), P3 major core (110 kDa), Pns9 nonstructural (83 kDa), P5 (89 kDa), Pns6 nonstructural (56 kDa), P7 minor core (58 kDa), P8 outercapsid (43 kDA), Pns9 nonstructural (49 kDa), Pns10 nonstructural (35 kDa), and Pns11a nonstructural (23 kDa) proteins. These molecular masses were in accord with those obtained from previous in vitro translation analysis. The locations of P2 and P5 remain to be determined although both of these are assumed to be outer layer proteins. Quantitative detection showed that accumulation (per gram of total proteins) of the virus-coded proteins in rice leaves is much greater (more than 15 times) than that in leafhoppers and that the content of the individual proteins varies within a sample from rice or leafhopper and also varies among different samples.

摘要

在杆状病毒载体系统中表达了12个水稻矮缩植物呼肠孤病毒(RDV)基因组片段中11个的截短型(S1和S2)或全长型(S3至S11)cDNA编码的肽段。针对每个表达肽段产生的抗体用作探针,以检测RDV开放阅读框编码的真实蛋白质。在RDV感染的水稻叶片和叶蝉(黑尾叶蝉)匀浆中被鉴定为S1至S11基因产物的多肽为P1小核心蛋白(170 kDa)、P2(130 kDa)、P3大核心蛋白(110 kDa)、Pns9非结构蛋白(83 kDa)、P5(89 kDa)、Pns6非结构蛋白(56 kDa)、P7小核心蛋白(58 kDa)、P8外衣壳蛋白(43 kDa)、Pns9非结构蛋白(49 kDa)、Pns10非结构蛋白(35 kDa)和Pns11a非结构蛋白(23 kDa)。这些分子量与先前体外翻译分析获得的分子量一致。尽管P2和P5都被认为是外层蛋白,但其定位仍有待确定。定量检测表明,水稻叶片中病毒编码蛋白的积累量(每克总蛋白)比叶蝉中的积累量高得多(超过15倍),并且单个蛋白的含量在来自水稻或叶蝉的样品内有所不同,在不同样品之间也有所不同。

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