TRANSGENIC AVIAN WHICH HAS FOREIGN GENE CONTAINING SEQUENCE ENCODING FELINE-DERIVED PROTEIN AND METHOD FOR PRODUCTION THEREOF
The present invention has for its object to provide a transgenic bird with a foreign gene containing a feline-derived protein-encoding sequence as transferred therein, and a method of producing the same. The present invention provides a method of producing a feline-derived protein by using a transgenic bird with a method which comprises infecting an avian embryo with a replication defective retrovirus vector containing a foreign gene by microinjection thereof into the early heart or blood vessel formed in the embryo and allowing the embryo to hatch.
1 . A transgenic bird
which has a foreign gene containing a sequence encoding a feline-derived protein.
2 . A transgenic bird
which has a foreign gene containing a sequence encoding a feline-derived cytokine protein and/or a protein substantially identical in biological activity thereto.
3 . A transgenic bird
which has a foreign gene containing at least a part of a sequence encoding the feline-derived erythropoietin identified under SEQ ID NO:1 in the sequence listing and/or a protein substantially identical in biological activity thereto.
4 . A transgenic bird
which has a foreign gene containing a sequence encoding the feline-derived erythropoietin identified under SEQ ID NO:1 in the sequence listing and/or a protein substantially identical in biological activity thereto.
5 . The transgenic bird according to claim 1 , which is produced by using a replication defective retrovirus vector.
6 . The transgenic bird according to claim 5 ,
wherein the replication defective retrovirus vector contains a sequence derived from Moloney murine leukemia virus and/or Moloney murine sarcoma virus.
7 . The transgenic bird according to claim 5 ,
wherein the replication defective retrovirus vector contains a sequence derived from murine stem cell virus.
8 . The transgenic bird according to claim 5 ,
wherein the replication defective retrovirus vector contains the VSV-G envelope sequence.
9 . The transgenic bird according to claim 5 ,
which is produced by using a replication defective retrovirus vector containing a non-tissue-specific promoter gene.
10 . The transgenic bird according to claim 9 ,
wherein the non-tissue-specific promoter gene contains a part or the whole of the chicken β-actin promoter gene.
11 . The transgenic bird according to claim 1 ,
which is produced by using a replication defective retrovirus vector containing a tissue-specific promoter gene.
12 . The transgenic bird according to claim 11 ,
wherein the tissue-specific promoter gene contains a part or the whole of an oviduct-specific promoter gene.
13 . The transgenic bird according to claim 12 ,
wherein the oviduct-specific promoter gene contains at least a part or the whole of the ovalbumin, ovotransferrin, ovomucoid, ovomutin, lysozyme, G2 globulin, G3 globulin, ovoinhibitor, ovoglycoprotein, ovoflavoprotein, ovomacroglobulin, cystatin or avidin promoter gene or a combination thereof.
14 . The transgenic bird according to claim 1 ,
which is produced by using a replication defective retrovirus vector containing a transcription enhancer and/or a regulatory element.
15 . The transgenic bird according to claim 14 ,
wherein the regulatory element contains a part or the whole of the woodchuck posttranscriptional regulatory element sequence.
16 . The transgenic bird according to claim 1 ,
which is obtained by a method comprising infecting an avian embryo with a replication defective retrovirus vector containing a foreign gene and allowing the embryo to hatch.
17 . The transgenic bird according to claim 16
which is obtained by a method comprising incubating a fertilized avian egg, infecting the embryo thereof after the lapse of at least 24 hours following the start of incubation with a replication defective retrovirus vector containing a foreign gene and allowing the embryo to hatch.
18 . The transgenic bird according to claim 17 ,
wherein the embryo is one formed during the period not earlier than 32 hours and not later than 72 hours after the start of incubation.
19 . The transgenic bird according to claim 18 ,
wherein the embryo is one formed during the period not earlier than 48 hours and not later than 64 hours after the start of incubation.
20 . The transgenic bird according to claim 16 ,
which is obtained by a method comprising infecting the embryo with the replication defective retrovirus vector by microinjection thereof into the heart or blood vessel formed in the embryo.
21 . The transgenic bird according to claim 1 ,
wherein said bird is a poultry bird.
22 . The transgenic bird according to claim 21 ,
wherein said bird is a chicken.
23 . The transgenic bird according to claim 1 , a descendant thereof, an egg thereof and spermatozoa thereof.
24 . A method of producing the transgenic bird according to claim 1 ,
which comprises infecting an avian embryo with a replication defective retrovirus vector containing a foreign gene and allowing the embryo to hatch.
25 . A method of producing a feline-derived protein
which comprises any or a combination of extracting the protein from the blood and/or somatic cells of the transgenic bird according to claim 1 and/or an egg laid thereby, purifying and activating the same.
26 . A polyethylene glycol-modified feline-derived protein
which is obtained by chemically modifying a feline-derived protein produced by the method according to claim 25 with polyethylene glycol.
27 . The polyethylene glycol-modified feline-derived protein according to claim 26 ,
wherein the feline-derived protein comprises a feline-derived cytokine protein and/or a protein substantially identical in biological activity thereto.
28 . The polyethylene glycol-modified feline-derived protein according to claim 26 ,
wherein the feline-derived protein is a protein containing at least a part of feline-derived erythropoietin identified under SEQ ID NO: 1 in the sequence listing and/or of a protein substantially identical in biological activity thereto.
29 . The polyethylene glycol-modified feline-derived protein according to claim 26 ,
wherein the feline-derived protein comprises feline-derived erythropoietin identified under SEQ ID NO: 1 in the sequence listing and/or a protein substantially identical in biological activity thereto.
30 . The polyethylene glycol-modified feline-derived protein according to claim 1 ,
wherein the polyethylene glycol used for the modification of the feline-derived protein has a weight average molecular weight of 5 to 40 kDa.
31 . The polyethylene glycol-modified feline-derived protein according to claim 30 ,
wherein the weight average molecular weight of the polyethylene glycol is 20 kDa.
32 . The polyethylene glycol-modified feline-derived protein according to claim 26 ,
wherein the number of polyethylene glycol molecules added is 1 or 2 or more and the apparent molecular weight, per polyethylene glycol-modified molecule, is from 100 kDa to 900 kDa as determined by gel filtration column chromatography in an aqueous solvent.
33 . The polyethylene glycol-modified feline-derived protein according to claim 32 ,
wherein the number of polyethylene glycol molecules added is 1 and the apparent molecular weight, per polyethylene glycol-modified molecule, is from 100 kDa to 500 kDa as determined by gel filtration column chromatography in an aqueous solvent.
34 . A polyethylene glycol-modified feline-derived protein composition
which comprises the polyethylene glycol-modified feline-derived protein according to claim 32 .
35 . A medicinal composition for feline use
which comprises, as an active ingredient, the polyethylene glycol-modified feline-derived protein according to claim 26 .
36 . A medicinal composition for feline use
which comprises, as an active ingredient, the polyethylene glycol-modified feline-derived protein composition according to claim 34 .
37 . The medicinal composition according to claim 35
which has feline erythropoietin activity and prolonged drug efficacy and is intended for use in the treatment of feline renal anemia.
38 . A method of producing the polyethylene glycol-modified feline-derived protein composition according to claim 34
which comprises causing a succinimidyl ester derivative of polyethylene glycol to add to a feline-derived protein.