Construction of recombinant myxoma viruses expressing foreign genes from different intergenic sites without associated attenuation Free

Abstract

Two myxoma virus transient dominant selection vectors were constructed and used to generate recombinant viruses expressing single and double foreign gene insertions from intergenic sites. The intergenic insertion sites were located between the myxoma virus genes MJ2 (thymidine kinase) and MJ2a, and MA24 (β-subunit RNA polymerase) and MA27 (fusion protein) located approximately 60 and 113 kb from the left-end of the viral genome, respectively. Recombinant myxoma viruses expressing the gene from either intergenic insertion site retained wild-type virulence. However, expression of the gene reduced the virulence of the recombinant viruses . Northern blot analysis indicated that the major late mRNAs encoding the viral RNA polymerase subunit and fusion protein are both of discrete size. Insertion of a foreign gene under the control of a synthetic late promoter between the MA24 and MA27 genes results in a specific-sized major late transcript for the inserted foreign gene. The MA27 gene transcripts directed by these recombinant viruses are heterogeneous in size, implying the typical pattern of poxvirus late transcription by random 3′-termination prior to polyadenylation. The transcription studies suggest signals located downstream of the insertion site direct 3′-processing of late transcripts irrespective of the gene immediately upstream.

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1996-07-01
2024-03-28
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References

  1. Amegadzie B. Y., Sister J. R., Moss B. 1992; Frame-shift mutations within the vaccinia virus A-type inclusion protein gene. Virology 186:777–782
    [Google Scholar]
  2. Antczak J. B., Patel D. D., Ray C. A., Ink B. S., Pickup D. J. 1992; Site-specific RNA cleavage generates the 3′ end of a poxvirus late mRNA. Proceedings of the National Academy of Sciences, USA 89:12033–12037
    [Google Scholar]
  3. Blasco R., Moss B. 1991; Extracellular vaccinia virus formation and cell-to-cell virus transmission are prevented by deletion of the gene encoding the 37000-dalton outer envelope protein. Journal of Virology 65:5910–5920
    [Google Scholar]
  4. Boyle D. B., Coupar B. E. H. 1988; A dominant selectable marker for construction of recombinant poxviruses. Gene 65:123–128
    [Google Scholar]
  5. Buller R. M. L., Smith G. L., Cremer K., Notkins A. L., Moss B. 1985; Decreased virulence of recombinant vaccinia virus expression vectors is associated with a thymidine kinase-negative phenotype. Nature 317:813–815
    [Google Scholar]
  6. Davison A. J., Moss B. 1989a; Structure of vaccinia virus early promoters. Journal of Molecular Biology 210:749–769
    [Google Scholar]
  7. Davison A. J., Moss B. 1989b; Structure of vaccinia virus late promoters. Journal of Molecular Biology 210:771–784
    [Google Scholar]
  8. Davison A. J., Moss B. 1990; New vaccinia virus recombination plasmids incorporating a synthetic late promoter for high level expression of foreign proteins. Nucleic Acids Research 18:4285–4286
    [Google Scholar]
  9. Falkner F. G., Moss B. 1990; Transient dominant selection of recombinant vaccinia viruses. Journal of Virology 64:3108–3111
    [Google Scholar]
  10. Fenner F., Marshall I. D. 1957; A comparison of the virulence for European rabbits (Oryctolagus cuniculus) of strains of myxoma virus recovered in the field in Australia, Europe and America. Journal of Hygiene 55:149–191
    [Google Scholar]
  11. Fenner F., Ratcliffe F. N. 1965 Myxomatosis Cambridge: Cambridge University Press;
    [Google Scholar]
  12. Gershon P. D., Black D. N. 1989a; The nucleotide sequence around the capripoxvirus thymidine kinase gene reveals a gene shared specifically with leporipoxvirus. Journal of General Virology 70:525–533
    [Google Scholar]
  13. Gershon P. D., Black D. N. 1989b; A capripoxvirus pseudogene whose only intact homologs are in other poxvirus genomes. Virology 172:350–354
    [Google Scholar]
  14. Gershon P. D., Ansell D. M., Black D. N. 1989; A comparison of the genome organization of capripoxvirus with that of the orthopoxviruses. Journal of Virology 63:4703–4708
    [Google Scholar]
  15. Goebel S. J., Johnson G. P., Perkus M. E., Davis S. W., Winslow J. P., Paoletti E. 1990; The complete DNA sequence of vaccinia virus. Virology 179:247–266
    [Google Scholar]
  16. Hall R. L., Moyer R. W. 1991; Identification, cloning, and sequencing of a fragment of Amsacta moorei entomopoxvirus DNA containing the spheroidin gene and three vaccinia virus–related open reading frames. Journal of Virology 65:6516–6527
    [Google Scholar]
  17. Herman G. E., O’Brien W. E., Beaudel A. L. 1986; An E. coli β-galactosidase cassette suitable for study of eukaryotic expression. Nucleic Acids Research 14:7130
    [Google Scholar]
  18. Holland M. K., Jackson R. J. 1994; Virus-vectored immunocontraception for control of wild rabbits: identification of target antigens and construction of recombinant viruses. Reproduction Fertility and Development 6:631–642
    [Google Scholar]
  19. Jackson R. J., Bults H. G. 1992a; The myxoma virus thymidine kinase gene: sequence and transcriptional mapping. Journal of General Virology 73:323–328
    [Google Scholar]
  20. Jackson R. J., Bults H. G. 1992b; A myxoma virus intergenic transient dominant selection vector. Journal of General Virology 73:3241–3245
    [Google Scholar]
  21. Kerr P. J., Jackson R. J. 1995; Myxoma virus as a vaccine vector for rabbits: antibody levels to influenza virus haemagglutinin presented by a recombinant myxoma virus. Vaccine 13:1722–1726
    [Google Scholar]
  22. Kochneva G. V., Urmanov I. H., Ryabchikova E. I., Streltsov V. V., Serpinsky O. I. 1994; Fine mechanisms of ectromelia virus thymidine kinase-negative mutants avirulence. Virus Research 34:49–61
    [Google Scholar]
  23. Meyer H., Osterrieder N., Czerny C. P. 1994; Identification of binding sites for neutralizing monoclonal antibodies on the 14-kDa fusion protein of orthopox viruses. Virology 200:778–783
    [Google Scholar]
  24. Massung R. F., Liu L.-I., Qi J., Knight J. C., Yuran T. E., Kerlavage A. R., Parsons J. M., Venter J. C., Esposito J. J. 1994; Analysis of the complete genome of smallpox variola major virus strain Bangladesh-1975. Virology 201:215–240
    [Google Scholar]
  25. Nasse M., Nicholson B. H., Fraser K. M., Mercer A. A., Robinson A. J. 1991; An orf virus sequence showing homology to the 14K ‘fusion’ protein of vaccinia virus. Journal of General Virology 72:1177–1181
    [Google Scholar]
  26. Opgenorth A., Graham K., Nation N., Strayer D., McFadden G. 1992; Deletion analysis of two tandemly arranged virulence genes in myxoma virus, MI1L and myxoma growth factor. Journal of Virology 66:4720–4731
    [Google Scholar]
  27. Patel D. D., Pickup D. J. 1987; Messenger RNAs of a strongly- expressed late gene of cowpox virus contain 5′-terminal poly(A) sequences. EMBO Journal 6:3787–3794
    [Google Scholar]
  28. Patel D. D., Pickup D. J. 1989; The second–largest subunit of the poxvirus RNA polymerase is similar to the corresponding subunits of procaryotic and eucaryotic RNA polymerases. Journal of Virology 63:1076–1086
    [Google Scholar]
  29. Pearson W. R., Lipman D. L. 1988; Improved tools for biological sequence comparison. Proceedings of the National Academy of Sciences, USA 85:2444–2448
    [Google Scholar]
  30. Rodriguez J. F., Smith G. L. 1990; IPTG-dependent vaccinia virus: identification of virus protein enabling virion envelopment by Golgi membrane and egress. Nucleic Acids Research 18:5347–5351
    [Google Scholar]
  31. Rodriguez D., Rodriguez J., Esteban M. 1993; The vaccinia virus 14-kilodalton fusion protein forms a stable complex with the processed protein encoded by the vaccinia virus A17L gene. Journal of Virology 67:3435–3440
    [Google Scholar]
  32. Rodriguez J. F., Paez E., Esteban M. 1987; A 14000-M, envelope protein of vaccinia virus is involved in cell fusion and forms covalently linked trimers. Journal of Virology 61:395–404
    [Google Scholar]
  33. Rosel J. L., Earl P. L., Weir J. P., 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 Hin dII H genome fragment. Journal of Virology 60:436–449
    [Google Scholar]
  34. Russell R. J., Robbins S. J. 1989; Cloning and molecular characterization of the myxoma virus genome. Virology 170:147–159
    [Google Scholar]
  35. Thompson J. D., Higgins D. G., Gibson T. J. 1994; CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, positions–specific gap penalties and weight matrix choice. Nucleic Acids Research 22:4673–4680
    [Google Scholar]
  36. Yuen L., Moss B. 1987; Oligonucleotide sequence signalling transcriptional termination of vaccinia virus early genes. Proceedings of the National Academy of Sciences, USA 84:6417–6421
    [Google Scholar]
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