Persistence of viral genes in a variant of MDBK cell after productive replication of a mutant of influenza virus A/WSN.
The MDBK-R cell line is a variant of the MDBK cell line, which was derived by three consecutive high multiplicity superinfections of MDBK cells with AWBY-140 virus, a mutant of influenza virus A/WSN (H 1N 1). MDBK-R cells are permissive for productive replication of AWBY-140, but resist lysis by the virus and grew normally without producing infectious virus after replication of the mutant occurred there. By polymerase chain reaction (PCR), we demonstrated nucleotide sequences specific to all the 8 genes of AWBY-140 in MDBK-R cells which had been infected with the mutant at a high multiplicity and subsequently received 25 passages. This suggests that the genes of influenza virus mutant persisted in the dividing host cells for a long time after productive infection, when none of the cells was producing virus. We were also able to amplify the M gene related sequence of the mutant from both poly(A)+ and poly(A)- fractions of the RNA extracted from the cells at 27th passage level by PCR, which suggests that the persisting genes were replicated and transcribed, but we failed to demonstrate any viral protein in the cells by Western blotting.
Urabe
M
M
Department of Virology, Jichi Medical School, Tochigi-Ken, Japan.
Tanaka
T
T
Odagiri
T
T
Tashiro
M
M
Tobita
K
K
D001483:Base Sequence; D002460:Cell Line; D004277:DNA, Single-Stranded; D004279:DNA, Viral / Q000032:analysis; D005814:Genes, Viral; D009980:Influenza A virus / Q000235:genetics* / Q000502:physiology; D008969:Molecular Sequence Data; D012692:Serial Passage; D014763:Viral Matrix Proteins / Q000235:genetics; D014779:Virus Replication
D004277:DNA, Single-Stranded; D004279:DNA, Viral; C031540:M-protein, influenza virus; D014763:Viral Matrix Proteins