Summary
Characterization of a late promoter of fowlpox virus (FPV) and a study of its activity in FPV and vaccinia virus (VV) was carried out. The 5′-mRNA start site of the FPV late gene mapped to a TAAAT sequence near the translation start site (ATG). A cloned DNA fragment of FPV genome (PFL1) comprising of the 5′-end of the late gene was used to express the LacZ gene ofE. coli in FPV and VV recombinants. A comparative analysis of β-galactosidase (BG) expression from the LacZ gene under the control of the FPV promoter and a VV late promoter (PL11) was performed. Like FPV-PL11-LacZ and VV-PL11-LacZ constructs, FPV-PFL1-LacZ and VV-PFL1-LacZ virus recombinants expressed BG indicating that essential features of transcription were conserved in the two viruses. Furthermore, the LacZ transcripts originating from PFL1 in FPV and VV recombinants mapped to the expected TAAAT sequence. Time course analysis of BG expressed by VV and FPV recombinants suggested that although the transcription machinery in the two viruses was essentially conserved, subtle differences in the efficiency of transcription or translation may exist.
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References
Binns MM, Boursnell MEG, Tomley FM, Campbell J (1989) Analysis of the fowlpoxvirus gene encoding the 4b core polypeptide and demonstration that it possesses efficient promoter sequences. Virology 170: 288–291
Binns MM, Stenzler L, Tomley FM, Campbell J, Boursnell MEG (1987) Identification by random sequencing strategy of the fowlpoxvirus DNA polymerase gene, its nucleotide sequence and comparison with other viral DNA polymerases. Nucleic Acids Res 15: 6563–6573
Boyle DB, Coupar BEH (1986) Identification and cloning of the fowlpox virus thymidine kinase gene using vaccinia virus. J Gen Virol 67: 1591–1600
Boyle DB, Coupar BEH (1988) Construction of recombinant fowlpox viruses as vectors for poultry vaccines. Virus Res 10: 343–356
Boyle DB, Coupar BEH (1988) A dominant selectable marker for the construction of recombinant poxviruses. Gene 65: 123–128
Boyle DB, Coupar BEH, Both G (1985) Multiple cloning site plasmids for the rapid construction of recombinant poxivurses. Gene 35: 169–177
Boyle DB, Coupar BEH, Gibbs AJ, Siegman LJ, Both G (1987) Fowlpox virus thymidine kinase: nucleotide sequence and relationships to other thymidine kinases. Virology 156: 355–365
Chakrabarti S, Brechling K, Moss B (1985) Vaccinia virus expression vector: coexpression of β-galactosidase provides visual screening of recombinant virus plaques. Mol Cell Biol 5: 3403–3409
Fenner F, Woodroofe GM (1960) The reactivation of poxviruses II. The range of reactivating viruses. Virology 11: 185–201
Hanggi M, Bannwarth W, Stunnenberg HG (1986) Conserved TAAAT motif in vaccinia virus late promoters: overlapping TATA box and site of transcription initiation. EMBO J 5: 1071–1076
Herman GE, O'Brien WE, Beaudet AL (1986) AnE. coli β-galactosidase cassette suitable for study of eukaryotic expression. Nucleic Acids Res 14: 7130
Kumar S, Boyle DB (1989) Mapping of a major early/late gene of fowlpox virus. Virus Res 15: 175–185
Lavin MF, Houldsworth J, Kumar S, Stein JL, Stein GS (1987) Coupling of histone mRNA levels to radioresistant DNA synthesis in ataxia-telangiectasia cells. Mol Cell Biochem 73: 45–54
Miller JH (1972) Experiments in molecular genetics. Cold Spring Harbor Laboratory, Cold Spring Harbor, New York
Moss B (1985) Replication of poxviruses. In: Fields BN (ed) Virology. Raven, New York, pp 685–703
Moss B, Flexner C (1987) Vaccinia virus expression vectors. Annu Rev Immunol 5: 305–324
Prideaux CT, Boyle DB (1987) Fowlpox virus polypeptides: sequential appearance and virion associated polypeptides. Arch Virol 96: 185–199
Prideaux CT, Kumar S, Boyle DB (1990) Comparative analysis of vaccinia virus promoter activity in fowlpox and vaccinia virus recombinants. Virus Res (In press)
Taylor J, Weinberg R, Languet B, Desmettre P, Paoletti E (1988) Recombinant fowlpox virus inducing protective immunity in non-avian species. Vaccine 6: 497–503
Taylor J, Weinberg R, Kawaoka Y, Webster RG, Paoletti E (1988) Protective immunity against avian influenza induced by a fowlpox virus recombinant. Vaccine 6: 504–508
Rosel JL, Earl PL, Weir JP, Moss B (1986) Conserved TAAATG sequence at the transcriptional and translational initiation sites of vaccinia virus late genes deduced by structural and functional analysis of the HindIII H genome fragment. J Virol 60: 436–449
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Kumar, S., Boyle, D.B. Activity of a fowlpox virus late gene promoter in vaccinia and fowlpox virus recombinants. Archives of Virology 112, 139–148 (1990). https://doi.org/10.1007/BF01323160
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DOI: https://doi.org/10.1007/BF01323160