Summary
In this report, the molecular weight and antigenicity of the proteins of a porcine enteric calicivirus (PEC) were characterized. The PEC virions were purified from intestinal contents of infected pigs and from infected cell culture lysates. The average buoyant density of the purified virus was 1.37 gm/cm3 in cesium chloride. One major structural protein with a molecular weight of approximately 58k was found in the gut and cell culture-passaged PEC using sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). Using immunoblotting techniques only one immunoreactive protein (58 k) ws identified. The PEC and a prototype calicivirus, feline calicivirus (FCV) were propagated in pig kidney and feline kidney (Crandell) cell lines, respectively and intrinsically labeled using [35S]methionine at various times post-inoculation (PI). SDS-PAGE of the radiolabeled proteins indicated the presence of the major structural protein (58 k) and one probable nonstructural protein (28 k) synthesized in the PEC-infected cell lysates by 12 h PI. Other minor protein bands were also evident by 24 h PI (32 k and 82 k). Only the 58 k major protein was detected by radioimmunoprecipitation (RIP) analysis using hyperimmune anti-PEC serum. SDS-PAGE and RIP analysis of FCV-infected cell lysates using hyperimmune anti-FCV serum identified a single major protein of approximately 64 k. No antigenic relationship between PEC and FCV proteins was detected by RIP analysis. The single major structural protein of PEC, the morphological appearance and size of the virus, and its average density of 1.37 gm/cm3 in cesium chloride are consistent with properties of other members of the familyCaliciviridae.
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References
Cubitt WD (1987) The candidate caliciviruses. In: Bock G, Whelan J (eds) Novel diarrhoea viruses. Wiley, Chichester, pp 126–143
Flynn WT, Saif LJ (1988) Serial propagation of porcine enteric calicivirus-like virus in primary porcine kidney cell cultures. J Clin Microbiol 26: 206–212
Flynn WT, Saif LJ, Moorhead PD (1988) Pathogenesis of porcine enteric calicivirus-like virus in four-day-old gnotobiotic pigs. Am J Vet Res 49: 819–925
Fretz M, Schaffer FL (1978) Calicivirus proteins in infected cells: evidence for a capsid polypeptide precursor. Virology 89: 318–321
Greenberg HB, Valdesuso J, Kalica AR, Wyatt RG, McAuliffe VJ, Kapikian AZ, Chanock RM (1981) Proteins of Norwalk virus. J Virol 37: 994–999
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of the bacteriophage T4. Nature 227: 680–685
Saif LJ, Bohl EH, Gupta RKP (1972) Isolation of porcine immunoglobulins and determination of the immunoglobulin classes of transmissible gastroenteritis viral antibodies. Infect Immun 6: 289–301
Saif LJ, Bohl EH, Kohler EM, Hughes JH (1977) Immune electron microscopy of transmissible gastroenteritis virus and rotavirus (reovirus-like agent) of swine. Am J Vet Res 38: 13–20
Saif LJ, Bohl EH, Theil KW, Cross RF, House JA (1980) Rotavirus-like, calicivirus-like, and 23-nm virus-like particles associated with diarrhea in young pigs. J Clin Microbiol 12: 105–111
Sammons DW, Adams LD, Nishizawa E (1981) Ultrasensitive silver based color staining of polypeptides in polyacrylamide gels. Electrophoresis 2: 135–141
Schaffer FL (1979) Caliciviruses: In: Fraenkel-Conrat H, Wagner RR (eds) Comprehensive virology, vol 14. Plenum, New York, pp 249–284
Schaffer FL, Bachrach HL, Brown F, Gillespie JH, Madin SH, Madeley CR, Povey RC, Scott F, Smith AW, Studdert MJ (1980) Caliciviridae. Intervirology 14: 1–6
Schaffer FL, Soergel ME, Black JW, Skilling DE, Smith AW, Cubitt WD (1985) Characterization of a new calicivirus isolated from the feces of a dog. Arch Virol 84: 181–195
Studdert MJ (1978) Caliciviruses. Arch Virol 58: 157–191
Terashima H, Chiba S, Sakuma Y (1983) The polypeptide of a human calicivirus. Arch Virol 78: 1–7
Welch SW, Saif LJ (1988) Monoclonal antibodies to a virulent strain of transmissible gastroenteritis virus: comparison of reactivity with virulent and attenuated virus. Arch Virol 101: 221–235
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Parwani, A.V., Saif, L.J. & Kang, SY. Biochemical characterization of porcine enteric calicivirus: analysis of structural and nonstructural viral proteins. Archives of Virology 112, 41–53 (1990). https://doi.org/10.1007/BF01348984
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DOI: https://doi.org/10.1007/BF01348984