Molecular cloning of the complete genome of reovirus serotype 3

Cashdollar, L W; Chmelo, Richard; Esparza, José; Hudson, G R; Joklik, Wolfgang K.

doi:10.1016/0042-6822(84)90438-0

Cited by 51 publications

(23 citation statements)

References 10 publications

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“…Two kinds of methods were used to clone rotavirus genes. The first kind was based on polyadenylation of genomic dsRNA, oligo(dT)-primed reverse transcription, followed by blunt-ended cloning or dC-tailing and cloning into dGtailored pBR322 (Imai et al, 1983;Cashdollar et al, 1984), or directly cloned into the TA vector pCR2.1 (Zhang & Rowhani, 2000), which is considered technically complicated, inefficient and also limited by the starting material (Potgieter et al, 2002). Other methods have been improved on the basis of a single-primer dsRNA cloning sequenceindependent method (Lambden et al, 1992).…”

Section: Discussionmentioning

confidence: 99%

Molecular characterization of a novel adult diarrhoea rotavirus strain J19 isolated in China and its significance for the evolution and origin of group B rotaviruses

Jiang

Tang

et al. 2008

Journal of General Virology

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The complete genome of a novel adult diarrhoea rotavirus strain J19 was cloned and sequenced using an improved single-primer sequence-independent method. The complete genome is 17 961 bp and is . Northern blot analysis and genomic sequence analysis indicated that segments 1-11 encode 11 viral proteins, respectively. Protein alignments with the corresponding proteins of J19 with B219, and groups A, B and C rotaviruses, produced higher per cent sequence identities to B219. Among groups A, B and C rotaviruses, 10 proteins from group B rotaviruses exhibited slightly higher amino acid sequence identity to the J19 proteins, but proteins of J19 showed low amino acid sequence identity with groups A and C rotaviruses. Construction of unrooted phylogenetic trees using a set of known proteins and representatives of three known rotavirus groups revealed that six structural proteins were positioned close to B219 and the basal nodes of groups A, B and C lineages, although with a preferred association with group B lineages. Phylogenetic analysis of the five non-structural proteins showed a similar trend. The results of the serological analysis, protein sequence analysis and phylogenetic analysis suggested that J19 would be a novel rotavirus strain with great significance to the evolution and origin of group B rotaviruses. INTRODUCTIONRotaviruses are important aetiological agents of disease in humans and animals. The viral genome is composed of 11 segments of double-stranded (ds) RNA that encode structural and non-structural proteins. The group A rotavirus strain SA11 genome, sequenced in 1990, is AUrich and has 18 555 bp (Both et al., 1984;Estes et al., 1984;Mitchell & Both, 1990). Its RNA segments encode six structural proteins (VP1, VP2, VP3, VP4, VP6 and VP7) and five non-structural proteins (NSP1, NSP2, NSP3, NSP4 and NSP5) (Estes, 2001). VP6 is the group antigen determinant, making up approximately 50 % of the viral protein (Estes et al., 1984;Estes & Cohen, 1989). The antigenic properties of VP6 are used to define seven antigenically different rotavirus groups, named groups A-G. Groups A, B and C rotaviruses infect humans, while other groups have been found only in animal species (Saif & Jiang, 1994). Group A rotaviruses cause severe diarrhoea in infants and young children, but they can also infect adults. Group C rotavirus infection occurs both in children and in adults, usually in sporadic cases or clustered outbreaks (Rodger et al., 1982;Caul et al., 1990;Jiang et al.,1995; Kuzuya et al., 1996; Nilsson et al., 2000;Adah et al., 2002;Chen et al., 2002;Ji et al., 2002;Schnagl et al., 2004).In China, large waterborne epidemics caused by the human group B rotavirus strain adult diarrhoea rotavirus (ADRV) infected thousands of people aged between 10 and 40 years of age in the 1980s (Hung et al., 1983(Hung et al., , 1984Chen et al., 1985Chen et al., , 1990 et al., 1987). The same rotavirus was implicated in gastroenteritis outbreaks in Beijing city in 1994 and in Shijiazhuang city in 1997. Partial genes of NADRV were clon...

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Section: Discussionmentioning

confidence: 99%

Molecular characterization of a novel adult diarrhoea rotavirus strain J19 isolated in China and its significance for the evolution and origin of group B rotaviruses

Jiang

Tang

et al. 2008

Journal of General Virology

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“…The type 3 Dearing (T3D) reovirus M3 genome segment was cloned by Cashdollar et al (7) and subcloned into pUC8 by Wiener et al (44). The clone was a generous gift from M. R. Roner (Florida Atlantic University).…”

Section: Methodsmentioning

confidence: 99%

Reovirus Nonstructural Protein μNS Binds to Core Particles but Does Not Inhibit Their Transcription and Capping Activities

Broering

McCutcheon

Centonze

et al. 2000

J Virol

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Previous studies provided evidence that nonstructural protein NS of mammalian reoviruses is present in particle assembly intermediates isolated from infected cells. Morgan and Zweerink (Virology 68: [455][456][457][458][459][460][461][462][463][464][465][466] 1975) showed that a subset of these intermediates, which can synthesize the viral plus strand RNA transcripts in vitro, comprise core-like particles plus large amounts of NS. Given the possible role of NS in particle assembly and/or transcription implied by those findings, we tested whether recombinant NS can bind to cores in vitro. The NS protein bound to cores, but not to two particle forms, virions and intermediate subvirion particles, that contain additional outer-capsid proteins. Incubating cores with increasing amounts of NS resulted in particle complexes of progressively decreasing buoyant density, approaching the density of protein alone when very large amounts of NS were bound. Thus, the NS-core interaction did not exhibit saturation or a defined stoichiometry. Negative-stain electron microscopy of the NS-bound cores revealed that the cores were intact and linked together in large complexes by an amorphous density, which we ascribe to NS. The NS-core complexes retained the capacity to synthesize the viral plus strand transcripts as well as the capacity to add methylated caps to the 5 ends of the transcripts. In vitro competition assays showed that mixing NS with cores greatly reduced the formation of recoated cores by stoichiometric binding of outer-capsid proteins 1 and 3. These findings are consistent with the presence of NS in transcriptase particles as described previously and suggest that, by binding to cores in the infected cell, NS may block or delay outer-capsid assembly and allow continued transcription by these particles.

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“…The double-stranded viral satellite RNAs peanut stunt virus associated RNA 5 (PARNA 5, 393 bp (18)) and cucumber mosaic virus associated RNA S (nCARNA 5, 335 bp (19)) were kindly supplied by J.Kaper. Genomic dsRNAs of reovirus Serotype 3 were a gift of H.Becht and H.Miuller; the 10 genomic segments of reovirus range from 1.2 to 3.8 kbp (20). L-dsRNA, the 4.3 kbp large genomic ragment of yeast killer factor (21) was prepared from the yeast strain YNN27.…”

Section: Materials and Methods Nucleic Acidsmentioning

confidence: 99%

Monoclonal antibodies to double-stranded RNA as probes of RNA structure in crude nucleic acid extracts

Schönborn¹,

Oberstrass²,

et al. 1991

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Molecular cloning of the complete genome of reovirus serotype 3

Cited by 51 publications

References 10 publications

Molecular characterization of a novel adult diarrhoea rotavirus strain J19 isolated in China and its significance for the evolution and origin of group B rotaviruses

Molecular characterization of a novel adult diarrhoea rotavirus strain J19 isolated in China and its significance for the evolution and origin of group B rotaviruses

Reovirus Nonstructural Protein μNS Binds to Core Particles but Does Not Inhibit Their Transcription and Capping Activities

Monoclonal antibodies to double-stranded RNA as probes of RNA structure in crude nucleic acid extracts

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