Purification and Preliminary Characterization of Infantile Gastroenteritis Virus (Orbivirus Group)

Petric, Martin; Szymanski, Maria T.; Middleton, Peter J.

doi:10.1159/000149919

Cited by 27 publications

(13 citation statements)

References 3 publications

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“…The pellets were resuspended in 4.0 ml of PBS and the virus was separated by equilibrium density centrifugation in cesium chloride (CsCI) as described by Petric ei at. [9] for the human rotavirus. The gradient fractions were dialyzed against PBS and then used in HA, HAI and CF tests.…”

Section: Methodsmentioning

confidence: 99%

Hemagglutination and Hemagglutination-inhibition Studies with a Strain of Nebraska Calf Diarrhea Virus (Bovine Rotavirus)

Fauvel¹,

Spence²,

Babiuk³

et al. 1978

Intervirology

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A hemagglutinin has been prepared from Nebraska calf diarrhea virus (NCDV) propagated in BS-C-1 cell line. After cesium chloride centrifugation, the hemagglutinin of the bovine rotavirus was found to be associated with intact virions (density 1.355 g/ml) but not with virions lacking an outer capsid layer (density 1.375 g/ml). In hemagglutination-inhibition (HAI) tests, the hemagglutinin reacted specifically with NCDV serum, and HAI seroconversions were detected in some sera tested. Cross-reactions were observed in complement fixation tests between the human and bovine rotaviruses but were not demonstrated by HAI, suggesting that the hemagglutinin detects a specific rather than a group antibody response.

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Section: Methodsmentioning

confidence: 99%

Hemagglutination and Hemagglutination-inhibition Studies with a Strain of Nebraska Calf Diarrhea Virus (Bovine Rotavirus)

Fauvel¹,

Spence²,

Babiuk³

et al. 1978

Intervirology

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show abstract

“…Two h after infection, the medium was changed to MEM containing 1 ~ of tl~e standard methionine concentration and at 4 h after infection [35S]methionine (New England Nuclear, 1274 Ci/mmol) was added to a final concentration of 10 gCi/ml. After an .additional 16 h of incubation at 37 °C the cells were harvested and the virus was purified by differential centrifugation, Freon extraction and CsCI gradient ultracentrifugation (Petric et al, 1975).…”

Section: Cells and Rirusmentioning

confidence: 99%

Oligosaccharide Composition of Calf Rotavirus

Kouvelos

Petric

Middleton

1984

Journal of General Virology

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SUMMARYThe oligosaccharides of the structural glycoprotein (VP7)of calf rotavirus were characterized. The precursor of VP7 produced in infected cells in the presence of tunicamycin migrated on SDS-polyacrylamide gels with an apparent molecular weight 6000 less than the glycosylated glycoprotein. Endoglycosidase (Endo) H digestion of the mature virus resulted in a decrease of 5000 in the molecular weight of VP7 in two discrete stages. Analysis of Endo H-treated, 3H-labelled digestion products of VP7 on Bio-Gel P-4 identified an oligosaccharide of molecular weight 1350 as the predominant form. Further treatment of the digest with mannosidase and analysis on Bio-Gel P-2 columns indicated that the oligosaccharide was digested into a free mannose and an oligosaccharide of molecular weight 400 in the ratio of 6:1. This indicates that the oligosaccharides of VP7 consist of four N-linked (Man)7 residues, two of which occupy more exposed and two more cryptic positions in the VP7 molecule.

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“…The virus was purified by subjecting the cells to a freeze-thaw cycle, followed by differential low-speed centrifugation. The cell pellet was extracted with Freon 113 and the aqueous phases subjected to two cycles of CsC1 isopycnic ultracentrifugation (Petric et al, 1975). The viruscontaining band of density 1.36 g/ml was further ultracentrifuged and the virus pellet resuspended in CP buffer (0.1 M-citric acid, 0.2 M-NazHPO4, pH 5.8).…”

mentioning

confidence: 99%

Comparison of Bovine, Simian and Human Rotavirus Structural Glycoproteins

Kouvelos¹,

Petric²,

Middleton³

1984

Journal of General Virology

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SUMMARYThe structural glycoproteins of calf (BDV 486), human (Wa) and Simian (SAIl) rotaviruses were compared for sensitivity to endoglycosidase (Endo) H. The calf and human virus glycoproteins were reduced by approximately 5000 molecular weight in two equal stages. The simian rotavirus glycoprotein sustained only one 2500 molecular weight decrease. The effect of the host cell on the viral oligosaccharide composition was examined. Endo H digests of calf rotavirus 82-124 extracted from stool and its progeny propagated and [35S]methionine-labelled in MA104 cells were compared. Endo H had the same effects on the glycoproteins, indicating that the nature of the host cell does not affect the oligosaccharide composition of the virus glycoproteins.

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Purification and Preliminary Characterization of Infantile Gastroenteritis Virus (Orbivirus Group)

Abstract: Infantile gastroenteritis virus (orbivirus group) was purified from stools of infants with acute viral diarrhea. The virus has a sedimentation coefficient of 520–530S and a buoyant density on CsCl of 1.36 g/ml.

Cited by 27 publications

References 3 publications

Hemagglutination and Hemagglutination-inhibition Studies with a Strain of Nebraska Calf Diarrhea Virus (Bovine Rotavirus)

Hemagglutination and Hemagglutination-inhibition Studies with a Strain of Nebraska Calf Diarrhea Virus (Bovine Rotavirus)

Oligosaccharide Composition of Calf Rotavirus

Comparison of Bovine, Simian and Human Rotavirus Structural Glycoproteins

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