Transient expression of B19 parvovirus gene products in COS-7 cells transfected with B19-SV40 hybrid vectors.
Hybrid B19 parvovirus-SV40 origin vectors were transfected into COS-7 cells and replication of these plasmids studied. Plasmids that have a frameshift mutation within the nonstructural gene region replicated to high level (copy number approximately 10,000/transfected cell) although somewhat lower than pSVOd, the SV40 origin vector without B19 sequence (copy number approximately 100,000/transfected cell). However, hybrid B19 parvovirus-SV40 origin vectors that do not contain these frameshift mutations replicated to a much lower level (copy number approximately 1000/transfected cell). Although the hybrid vectors studied replicated at different efficiencies in COS-7 cells, they are transcribed at approximately the same level, resulting in RNA species that are indistinguishable from those seen in B19 virus-infected erythroid bone marrow cells. Western blot analysis demonstrated that the mRNAs are translated into polypeptides of the same size and, in the case of viral structural proteins, in same relative abundance as seen in a B19-infected clinical sample.
Beard
C
C
Department of Biochemistry, Faculty of Medicine, University of British Columbia, Vancouver, Canada.
St Amand
J
J
Astell
C R
CR
D015152:Blotting, Northern; D015153:Blotting, Western; D002460:Cell Line; D004261:DNA Replication; D004279:DNA, Viral / Q000096:biosynthesis / Q000235:genetics; D015870:Gene Expression; D005822:Genetic Vectors; D006801:Humans; D010321:Parvoviridae / Q000235:genetics* / Q000502:physiology; D010957:Plasmids; D012326:RNA Splicing; D012367:RNA, Viral / Q000235:genetics; D014158:Transcription, Genetic; D014162:Transfection; D014764:Viral Proteins / Q000096:biosynthesis / Q000235:genetics; D014779:Virus Replication
D004279:DNA, Viral; D012367:RNA, Viral; D014764:Viral Proteins