EXPRESSION, CHARACTERIZATION, AND PURIFICATION OF A PHOSPHORYLATED RABIES NUCLEOPROTEIN SYNTHESIZED IN INSECT CELLS BY BACULOVIRUS VECTORS

被引：39

作者：

PREHAUD, C ^{[1
]}

HARRIS, RD ^{[1
]}

FULOP, V ^{[1
]}

KOH, CL ^{[1
]}

WONG, J ^{[1
]}

FLAMAND, A ^{[1
]}

BISHOP, DHL ^{[1
]}

机构：

[1] CNRS,GENET VIRUS LAB,F-91198 GIF SUR YVETTE,FRANCE

来源：

VIROLOGY | 1990年 / 178卷 / 02期

关键词：

D O I：

10.1016/0042-6822(90)90346-S

中图分类号：

Q93 [微生物学];

学科分类号：

071005 ; 100705 ;

摘要：

A baculovirus expression vector (AcNPV3) derived from the nuclear polyhedrosis virus of Autographa californica (AcNPV) was prepared containing the complete coding region of the nucleoprotein (N) gene of rabies virus (Gif-sur-Yvette clone of the CVS strain). The gene was placed under the control of the AcNPV polyhedrin promoter and was expressed to high levels (66 mg N protein/liter of 2 × 109 cells) by the derived recombinant virus using a Spodoptera frugiperda cell line. Using available antisera, it was established that the antigenic characteristics of the N protein were similar by comparison with those of the native N protein of rabies virus. Characterization of the expressed protein established that, like the N protein of mammalian cell-grown CVS virus, the N protein was phosphorylated. The expressed rabies N protein induced antibodies in mice that reacted strongly with the rabies viral protein. The expressed nucleoprotein was recovered from the insect cells by differential centrifugation followed by ion exchange chromatography. The expressed rabies N protein represents a source of authentic protein suitable for virus diagnosis as well as structural studies. © 1990.

引用

页码：486 / 497

页数：12

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[11] USE OF HYBRIDOMA MONOCLONAL-ANTIBODIES IN THE DETECTION OF ANTIGENIC DIFFERENCES BETWEEN RABIES AND RABIES-RELATED VIRUS PROTEINS .1. THE NUCLEOCAPSID PROTEIN [J].