Recombinant Avipox virus encoding polypeptide of mycoplasma gall

Drug – bio-affecting and body treating compositions – Antigen – epitope – or other immunospecific immunoeffector – Recombinant virus encoding one or more heterologous proteins...

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4242641, 4242141, 4242151, 4241901, 4242321, 4241921, 435 691, 435 693, 435 701, 4353201, 536 234, 536 237, 530350, 530820, 530825, 930200, A61K 39295, A61K 3912, C12N 1531, C07K 701

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058717420

DESCRIPTION:

BRIEF SUMMARY
This is a rule 371 application based on the Priority date of PCT/JP99/00541 filed Mar. 31, 1994.


TECHNICAL FIELD

The present invention relates to a novel polypeptide showing antigenicity to Mycoplasma gallisepticum, a fused polypeptide between the said polypeptide and a signal membrane anchor, and a recombinant Avipox virus capable of expressing a polypeptide showing antigenicity to Mycoplasma gallisepticum, especially a polypeptide showing antigenicity on the membrane surface of a host cell, as well as use thereof.


BACKGROUND

It is expected that a polypeptide showing antigenicity to Mycoplasma gallisepticum can be utilized as an effective ingredient of a vaccine for Mycoplasma gallisepticum infections, since an egg-laying rate and a hatching rate of eggs produced by infected chickens are markedly reduced when infected with Mycoplasma gallisepticum. At present, the system using Escherichia coli or yeast is known to prepare the antigenic protein of Mycoplasma gallisepticum by genetic engineering (Japanese Patent Application Laid-Open No. 2-111795). In general, it is pointed out that the production of a polypeptide in the system using bacteria involves problems that firstly an antigen is expressed in a less amount and secondly, a pyrogen originating in a host cannot be removed. It is thus the actual situation that such a system has not been practically applied yet. For this reason, studies have been made on the preparation of a polypeptide expressing an antigenicity or a recombinant live vaccine, using a recombinant virus. However, as far as Mycoplasma gallisepticum is concerned, any recombinant virus inserted with DNA encoding said protein has not been prepared.
In a virus protein where the virus infects cells, one type of a protein expressed is transported to the cell surface and the protein is expressed on the surface of a cell membrane (hereinafter such a state is sometimes merely referred to as being expressed on the cell surface) and another type of a protein that is not expressed on the cell surface. A representative example of the former protein is a glycoprotein contained in the coat of a virus. A recombinant virus that expresses such a protein efficiently exhibits the protein on the cell surface. It is thus considered that a high antibody titer can be induced in poultry infected with this recombinant virus (Japanese Patent Application Laid-Open No. 1-157381). On the other hand, an example of the latter type of protein includes a protein originating in bacteria, such as an antigenic protein of Mycoplasma gallisepticum.
It is not expectable to induce a high antibody titer from such recombinant viruses that express these proteins, since they are expressed on the cell membrane surface merely in an extremely small quantity. However, if such a protein can be expressed on the cell membrane surface in a large quantity by genetic engineering, a high antibody titer will be induced. Thus, investigations have been made to express on the membrane surface such a protein that is not principally expressed on the membrane surface. For example, there is a report that DNA encoding a signal protein having the function of secreting a protein on the cell membrane surface and DNA encoding a membrane anchor protein having the function of retaining the secreted protein so as not to leave out of the cell membrane surface are ligated with the 5' end and the 3' end of DNA encoding an antigenic protein, respectively, and a recombinant vaccinia virus inserted with the resulting hybrid DNA expresses the antigenic protein on the cell membrane surface of a host (J. Viol., 64, 4776-4783 (1990) or Mol. Cell. Biol., 6, 3191-3199 (1986)). However, DNA encoding a signal and DNA encoding a membrane anchor are independently ligated with DNA encoding an antigenic protein in these examples so that it is hardly applicable practically due to complicated preparation of a recombinant virus.


DISCLOSURE OF THE INVENTION

The present inventors have made extensive studies to provide a polypeptide having antigenicity originating in Mycoplas

REFERENCES:
patent: 5093258 (1992-03-01), Cohen et al.
patent: 5196514 (1993-03-01), Avakian et al.
Avakian et al, "Evaluation of Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis Purified Proteins of Mycoplasma gallisepticum and M. synoviae as Antigens in a Dot-Enzyme-Linked Immunosorbent Assay", Avian Diseases, vol. 34, pp. 575-584, 1990.
Molecular and Cellular Biology, vol. 10, No. 2, (1990), Wilson C. et al.: "Abenant membrane insertion of a cytoplasmic tail delection mutant of the hemagglutinin neuraminidase glycoprotein of newcastle disease virus", see pp. 449-457.
WO, A, 9324646 (Nippon Zeon Co., Ltd., Shionogi & Co., Ltd.), Dec. 9, 1993 (Sep. 12,1993) & AU, A, 9340903.

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