Chemistry: molecular biology and microbiology – Vector – per se
Patent
1991-03-20
1994-12-20
Nucker, Christine M.
Chemistry: molecular biology and microbiology
Vector, per se
536 241, 536 2372, 4352351, 935 6, 935 32, C12N 1579, C12N 1586, C12N 1511, C12N 1539
Patent
active
053745582
DESCRIPTION:
BRIEF SUMMARY
BACKGROUND OF THE INVENTION
1. Field of the Invention
The invention is in the field of recombinant DNA technology and relates to an avipox, especially fowlpox, virus promoter useful for the expression of foreign DNA inserted Into a fowlpox virus vector.
2. Description of the Prior Art
Poxviruses are large viruses with a complex morphology containing linear double-stranded DNA genomes. They are among the few groups of DNA viruses that replicate within the cytoplasm of the cell. They are subclassified into six genera: orthopoxviruses, avipoxviruses, capripoxviruses, leporipoxviruses, parapoxviruses and entomopoxviruses. Vaccinia virus (VV), an orthopoxvirus, is the most widely studied of the poxviruses, and is the subject of U.S. Pat. No. 4,603,112 (Paoletti et al.,). Fowlpox virus (FPV) is an avipoxvirus or avian poxvirus.
Recent advances in recombinant DNA technology have allowed VV to be used as a vector to carry and express foreign genes. Foreign DNA Is Introduced Into the VV genome by a process of homologous recombination. Homologous recombination involves essentially (1) pre-selecting a length of the VV genome in some region which does not impair the replication and normal functioning of the virus (hereinafter called a "non-essential region"), (2) making a construct comprising a VV promoter and a length of foreign DNA within a copy of the non-essential region (NER) so that the foreign DNA is under the control of the promoter and so that the promoter-foreign DNA combination is flanked by extensive sequences of non-essential region of VV DNA, (3) co-infecting appropriate tissue culture cells with the VV and with the construct and (4) selecting cells containing VV in which the pre-selected length has been recombined in vivo so that it is replaced in the genome by the construct DNA. The recombinant W expresses the foreign gene in vivo, stimulating the immunity to the protein in an appropriate host. The procedure has considerable potential for use in vaccination.
More recently, similar technology has been applied to fowlpox virus (FPV). Although VV promoters have been used successfully in laboratory constructs of FPV, It is undesirable to incorporate elements of such VV, an orthopoxvirus which has a wide host range recombinant vaccine, for fear of recombination events which could pose a health risk. There is therefore a need to develop FPV promoters for use in recombinant FPV. Certain FPV promoters designated as promoters of the "FP4b" gene have been described in UK Patent Application 8824746, now Publication No. 2211504A, or PCT Application GB 88/00922, now Publication No. W0/89/03879 (NRDC). However, each promoter has its own peculiar characteristics of strength and timing of promotion. A choice of promoters is therefore very highly desirable.
SUMMARY OF THE INVENTION
When the prior applications were filed, both on Oct. 21, 1988, their subject matter was enlarged shortly before filing to include the promoter of another gene known as the "790 bp gene" because it lay within a 790 base pair EcoRI fragment of the strain of FPV under investigation. The two prior applications were our first applications in any country for this particular subject matter. As a result of further work, it is now possible to define the "790 bp gene", and therefore the promoter, more precisely. The present invention provides this promoter and this application claims priority of UKPA 8824746 filed Oct. 21, 1988 and of another application filed on Apr. 21, 1989, before any publication of the promoter occurred in the prior applications (or elsewhere).
The promoter of the invention is defined by reference to the gene ORF 1 identified in the sequence of the 790 bp fragment shown below. Note that this sequence is provided in the other strand to that shown in UKPA 8824746, but is arranged in the same 5' and 3' direction. Further, the exact length of the fragment has been determined as 783 bp. Consequently, position 1 in the sequence below Is the complement of position "795" (an arbitrary number) in UKPA 8824746, while position 758 in th
REFERENCES:
patent: 5093258 (1992-03-01), Cohen et al.
patent: 5174993 (1992-12-01), Paoletti
Weir, J. P. et al. 1984, J. Virol. vol. 51 pp. 662-669.
Coupar, B. E. H. et al. 1986. Eur. J. Immunol. vol. 16 pp. 1479-1487.
Prideaux, C. T. et al. 1987. Arch. Virol. vol. 96 pp. 185-199.
Binns Matthew M.
Boursnell Michael E. G.
Campbell Joan I. A.
British Technology Group Ltd.
Mosher Mary E.
Nucker Christine M.
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