Claims for Patent: 6,548,735
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Summary for Patent: 6,548,735
Title: | Production of heterologous protein in milk of transgenic non-human mammals comprising a construct with an intron |
Abstract: | Proteinaceous products can be produced by transgenic animals having genetic constructs integrated into their genome. The construct comprises a 5\'-flanking sequence from a mammalian milk protein gene (such as beta-lactoglobulin) and DNA coding for a heterologous protein other than the milk protein (for example a serin protease such as alpha.sub.1 -antitrypsin or a blood factor such as Factor VIII or IX). The protein-coding DNA comprises at least one, but not all, of the introns naturally occurring in a gene coding for the heterologous protein. The 5\'-flanking sequence is sufficient to drive expression of the heterologous protein. |
Inventor(s): | Archibald; Alan Langskill (Edinburgh, GB), Clark; Anthony John (Midlothian, GB) |
Assignee: | PPL (Holdings) Limited (London, GB) |
Application Number: | 08/781,518 |
Patent Claims: | 1. A method for producing a heterologous protein, the method comprising expressing said heterologous protein in the milk of a non-human transgenic placental mammal and
harvesting the heterologous protein, wherein said non-human transgenic placental mammal has integrated into its genome a genetic construct comprising (a) a 5'-flanking sequence from a mammalian milk protein gene and (b) DNA coding for said heterologous
protein other than said milk protein, wherein said DNA comprises at least one whole, but not all, of the introns naturally occurring in the gene coding for said heterologous protein and wherein said 5'-flanking sequence controls expression of said
heterologous protein, said genetic construct being integrated into said non-human transgenic placental mammal's genome in such a way that said DNA is expressed in a mammary gland of said mammal at levels greater than those achieved by expression of a
corresponding intron-less construct.
2. A method as claimed in claim 1, wherein the transgenic non-human placental mammal transmits the construct to its progeny. 3. A method as claimed in claim 1, wherein the 5'-flanking sequence includes a P-lactoglobulin transcription start site consisting of the SphI-BglII 800 base pair fragment of pSS1tgSpDELTA-AvaII, or any fragment of said SphI-BglII 800 base pair fragment of pSS1tgSpDELTA-AvaII that contains said .beta.-lactoglobulin transcription start site. 4. A method as claimed in claim 1, wherein the 5'-flanking sequence includes a .beta.-lactoglobulin transcription start site consisting of the 4.2 kilobase pair SalI-SphI DNA fragment isolated from the 5'-flanking sequences of the .beta.-lactoglobulin gene or any fragment of said 4.2 kilobase pair SalI-SphI DNA fragment that contains said .beta.-lactoglobulin transcription start site. 5. A method as claimed in claim 1, wherein the milk protein gene is a .beta.-lactoglobulin gene. 6. A method as claimed in claim 1, wherein the heterologous protein is a serine protease inhibitor. 7. A method as claimed in claim 1, wherein the heterologous protein is a blood factor. 8. A method as claimed in claim 1, in which all but one of the natural introns is present in said DNA. 9. A method as claimed in claim 1, in which only one of the natural introns is present in said DNA. 10. A method as claimed in claim 1, in which the construct comprises a 3'flanking region derived from said mammalian milk protein gene or from said DNA coding for said heterologous protein. |
Details for Patent 6,548,735
Applicant | Tradename | Biologic Ingredient | Dosage Form | BLA | Approval Date | Patent No. | Expiredate |
---|---|---|---|---|---|---|---|
Merck Sharp & Dohme Corp. | INTRON A | interferon alfa-2b | For Injection | 103132 | 06/04/1986 | ⤷ Try a Trial | 2008-11-11 |
Merck Sharp & Dohme Corp. | INTRON A | interferon alfa-2b | For Injection | 103132 | ⤷ Try a Trial | 2008-11-11 | |
Merck Sharp & Dohme Corp. | INTRON A | interferon alfa-2b | Injection | 103132 | ⤷ Try a Trial | 2008-11-11 | |
>Applicant | >Tradename | >Biologic Ingredient | >Dosage Form | >BLA | >Approval Date | >Patent No. | >Expiredate |
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