An infectious cDNA copy of the genome of a non-cardiovirulent coxsackievirus B3 strain: its complete sequence analysis and comparison to the genomes of cardiovirulent coxsackieviruses.
The genome of the non-cardiovirulent coxsackievirus B3 (CVB3) strain CVB3/0 was cloned and sequenced to aid in the elucidation of the viral genetic basis for the CVB3 cardiovirulent phenotype. Reverse-transcribed sub-genomic complementary DNA (cDNA) fragments were enzymatically amplified using generic oligonucleotide primers and were assembled as a complete infectious genomic copy (pCVB3-0) downstream of the T7 RNA polymerase promoter. Positive-strand viral RNA transcribed from pCVB3-0 using T7 RNA polymerase and transfected into HeLa cells produced infectious virus (CVB3/0c). No differences in phenotype were observed comparing growth of CVB3/0c to the parental CVB3/0 in HeLa single-step growth curves, virus yields, or plaque size. When inoculated into C3H/HeJ mice, CVB3/0c achieved cardiac titers equivalent to the parental CVB3/0 and like the parental virus, demonstrated a non-cardiovirulent phenotype. The nucleotide sequence of the cloned CVB3/0 genome was determined and compared to the genomes of infectious cDNA clones of cardiovirulent CVB3 strains. Two consistent differences among nucleotides in non-translated regions and 8 amino acid differences relative to two well-characterized infectious cDNA copies of genomes from cardiovirulent CVB3 strains were identified.
Chapman
N M
NM
Department of Pathology and Microbiology, University of Nebraska Medical Center, Omaha.
Tu
Z
Z
Tracy
S
S
Gauntt
C J
CJ
D000818:Animals; D001483:Base Sequence; D002213:Capsid / Q000235:genetics; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary / Q000032:analysis; D004279:DNA, Viral / Q000737:chemistry / Q000235:genetics; D029822:Enterovirus B, Human / Q000235:genetics* / Q000472:pathogenicity* / Q000502:physiology; D016679:Genome, Viral; D006367:HeLa Cells; D006321:Heart / Q000382:microbiology*; D006801:Humans; D008297:Male; D051379:Mice; D008809:Mice, Inbred C3H; D008969:Molecular Sequence Data; D015183:Restriction Mapping; D013045:Species Specificity; D014162:Transfection; D017361:Viral Nonstructural Proteins / Q000235:genetics; D014779:Virus Replication
D017931:DNA Primers; D018076:DNA, Complementary; D004279:DNA, Viral; D017361:Viral Nonstructural Proteins