Detection of Rice Tungro Bacilliform Virus Gene Products in Vivo

• Published in: Virology (New York, N.Y.) Vol. 205; no. 2; pp. 430 - 437
• Main Authors: Hay, Joanne, Grieco, Francesco, Druka, Arnis, Pinner, Marion, Lee, Sor-Cheng, Hull, Roger
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.12.1994
• Subjects: Aspartic Acid Endopeptidases - genetics; Base Sequence; Blotting, Western; Escherichia coli; EXPRESION GENICA; EXPRESSION DES GENES; Immune Sera; IMMUNOLOGIE; INMUNOLOGIA; Microscopy, Immunoelectron; Molecular Sequence Data; Open Reading Frames; Oryza - virology; ORYZA SATIVA; Plant Diseases; Plant Viruses - enzymology; Plant Viruses - genetics; Polymerase Chain Reaction; PROTEINAS; PROTEINE; Recombinant Fusion Proteins - biosynthesis; Recombinant Fusion Proteins - isolation & purification; Ribonuclease H - genetics; rice tungro bacilliform virus; RNA-Directed DNA Polymerase - genetics; SECUENCIA NUCLEICA; SEQUENCE NUCLEIQUE; Viral Proteins - biosynthesis; Viral Proteins - genetics; Viral Proteins - isolation & purification; VIRUS DE LAS PLANTAS; VIRUS DEL TUNGRO DEL ARROZ; VIRUS DES VEGETAUX; VIRUS TUNGRO DU RIZ
• ISSN: 0042-6822; 1096-0341
• DOI: 10.1006/viro.1994.1663

To study the products of the open reading frames (ORFs) of rice tungro bacilliform virus in rice plants the sequences containing ORFs I (encoding a 24-kDa protein, P24) and IV (P46) and the protease and polymerase (reverse transcriptase + RNaseH) domains of ORF III were cloned into a pGEX expression vector. The proteins, which were C-terminal fusions to glutathione S-transferase, were expressed in Escherichia coli and antisera were raised against them which, together with an antiserum against virus particles, was used to probe blots of proteins from infected and uninoculate plants and from virus preparations. The P24 antiserum detected virus-specific proteins of 74, 60, and 52 kDa, which are much bigger than expected. These proteins were found in virus preparations and immunogold labeling suggested that they might be internal in the particles. Virus-specific proteins of 33, 37, 62, and >150 KDa were revealed by antiserum to virus particles. The antiserum to the protease revealed proteins of 13.5, 37, and 68 kDa both in extracts from infected plants and in purified virus preparations. This antiserum decorated intact virus particles as did the particle antiserum. The polymerase domain antiserum reacted with products of 56, 65, and 68 kDa in extracts from infected plants but not in virus particles. The antiserum to the ORF IV product did not detect any bands in either infected plant extracts or virus preparations. The significance of these products is discussed.