Claims for Patent: 6,214,578

Summary for Patent: 6,214,578

Title:	Method for the expressing foreign genes and vectors therefor
Abstract:	A method for expressing a foreign gene, by which the foreign genes can be expressed more strongly than by the conventional methods, as well as a recombinant vector therefor, is disclosed. In the method for expressing a foreign gene according to the present invention, the foreign gene is inserted into a site downstream of a promoter and the foreign gene is expressed in a cell, wherein a plurality of intron-originated DNA fragments which are the same or different and are capable of promoting expression of foreign genes are inserted into one or more sites upstream of the foreign gene, and said foreign gene is expressed.
Inventor(s):	Ueki; Jun (Shizuoka, JP), Ohta; Shozo (Shizuoka, JP), Morioka; Shinji (Tokyo, JP), Kuraya; Yoshiki (Shizuoka, JP)
Assignee:	Japan Tobacco Inc. (Tokyo, JP)
Application Number:	09/011,526
Patent Claims:	1. A method for expressing a foreign gene comprising: inserting said foreign gene into a site downstream of a promoter; and expressing said foreign gene in a cell, wherein a plurality of intron-originated DNA fragments selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, a variant having not less than 90% homology to SEQ ID NO:1, a variant having not less than 90% homology to SEQ ID NO:2, and a variant having not less than 90% homology to SEQ ID NO:3, which are the same or different and promote expression of foreign genes, are inserted into one or more sites upstream of said foreign gene; and wherein the level of expression of said foreign gene is higher than when only one of said intron-originated DNA fragments is inserted into a site upstream of said foreign gene. 2. The method according to claim 1, wherein said plurality of intron-originated DNA fragments are inserted into one or more sites between said promoter and said foreign gene. 3. The method according to claim 1 or 2, wherein said cell is a plant cell. 4. The method of claim 1, wherein at least one of said plurality of intron-originated DNA fragments consists of the sequence shown in SEQ ID NO;1 or a functional variant thereof having not less than 90% homology to SEQ ID NO:1. 5. The method according to claim 4, wherein said plurality of intron-originated DNA fragments comprises at least two copies consisting of the sequence shown in SEQ ID NO:1 or a functional variant thereof having not less than 90% homology to SEQ ID No:1. 6. The method according to claim 1, wherein at least one of said plurality of intron-originated DNA fragments consists of the sequence shown in SEQ ID No:2 or a functional variant thereof having not less than 90% homology to SEQ ID NO:2. 7. The method according to claim 6, wherein said plurality of intron-originated DNA fragments consists of the sequence shown in SEQ ID NO:1 or a functional variant thereof having not less than 90% homology to SEQ ID NO:1 and the sequence shown in SEQ ID NO:2 or a functional variant thereof having not less than 90% homology to SEQ ID NO:2. 8. The method according to claim 1, wherein at least one of said plurality of intron-originated DNA fragments consists of the sequence shown in SEQ ID NC:3 or a functional variant thereof having not less than 90% homology to SEQ ID NO:3. 9. The method according to claim 8, wherein said plurality of intron-originated fragments consists of the sequence shown in SEQ ID NO:1 or a functional variant thereof having not less than 90% homology to SEQ ID NO:1 and the sequence shown in SEQ ID NO:3 or a functional variant thereof having not less than 90% homology to SEQ ID NO:3. 10. A recombinant vector comprising: a promoter; a foreign gene inserted into a site downstream of said promoter; and a plurality of intron-originated DNA fragments which are the same or different and which promote expression of foreign genes, which are inserted into one or more sites upstream of said foreign gene, wherein said intron-originated DNA fragments consists of a sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3 and a sequence having not less than 90% homology to one of SEQ ID NO:1, SEQ ID NO:2 or SEQ ID NO:3. 11. The recombinant vector according to claim 10, wherein said plurality of intron-originated DNA fragments are inserted into one or more sites between said promoter and said foreign gene. 12. The recombinant vector according to claim 10 or 11, which are replicable in a plant cell. 13. The recombinant vector according to claim 10, wherein at least one of said plurality of intron-originated DNA fragments consists of the sequence shown in SEQ ID NO;1 or a functional variant thereof having not less than 90% homology to SEQ ID NO:1. 14. The recombinant vector according to claim 13, wherein said plurality of intron-originated DNA fragments comprises at least two copies consisting of the sequence shown in SEQ ID NO:1 or a functional variant thereof having not less than 90% homology to SEQ ID NO:1. 15. The recombinant vector according to claim 10, wherein at least one of said plurality of intron-originated DNA fragments consists of the sequence shown in SEQ ID NO:2 or a functional variant thereof having not less than 90% homology to SEQ ID NO:2. 16. The recombinant vector according to claim 15, wherein said plurality of intron-originated DNA fragments consists of the sequence shown in SEQ ID NO:1 or a functional variant thereof having not less than 90% homology to SEQ ID NO:1 and the sequence shown in SEQ ID NO:2 or a functional variant thereof having not less than 90% homology to SEQ ID NO:2. 17. The recombinant vector according to claim 10, wherein at least one of said plurality of intron-originated DNA fragments consists of the sequence shown in SEQ ID NO:3 or a functional variant thereof having not less than 90% homology to SEQ ID NO;3. 18. The recombinant vector according to claim 17, wherein said plurality of intron-originated DNA fragments consists of the sequence shown in SEQ ID NO:1 or a functional variant thereof having not less than 90% homology to SEQ ID NO:1 and the sequence shown in SEQ ID NO:3 or a functional variant thereof having not less than 90% homology to SEQ ID NO:3. 19. A method for expressing a foreign gene, comprising: inserting said foreign gene into a site downstream of a promoter; and expressing said foreign gene in a cell, characterized in that the sequence shown in SEQ ID NO:3 or a functional variant thereof having not less than 90% homology to SEQ ID NO:3 is inserted into a site upstream of said foreign gene, and said foreign gene is expressed. 20. A method for expressing a foreign gene, comprising: inserting said foreign gene into a site downstream of a promoter; and expressing said foreign gene in a cell, wherein the sequence shown in SEQ ID NO:3 is inserted into a site upstream of said foreign gene, and said foreign gene is expressed. 21. The method according to claim 19, wherein the sequence shown in SEQ ID NO:3 or a functional variant thereof having not less than 90% homology to SEQ ID NO:3 is inserted between said promoter and said foreign gene. 22. The method according to claim 19, wherein said cell is a plant cell. 23. The method of claim 20, wherein said sequence shown in SEQ ID NO:3 is inserted between said promoter and said foreign gene. 24. The recombinant vector according to claim 10, wherein said at least one of said plurality of intron-originated DNA fragments consists of a sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3. 25. A recombinant vector comprising: a promoter; a foreign gene inserted into a site downstream of said promoter; and a plurality of intron-originated DNA fragments which are the same or different and which promote expression of foreign genes, which are inserted into one or more sites upstream of said foreign gene, wherein each of said plurality of intron-originated DNA fragments consists of a sequence selected from the group consisting of SEQ ID NO:1, SEQ ID NO:2, and SEQ ID NO:3. 26. The recombinant vector according to claim 15, wherein said plurality of intron-originated DNA fragments consists of the sequence shown in SEQ ID NO:2 or a functional variant thereof having not less than 90% homology to SEQ ID NO:2 and the sequence shown in SEQ ID NO:3 or a functional variant thereof having not less than 90% homology to SEQ ID NO:3. 27. The recombinant vector according to claim 10, wherein expression of said foreign gene upon introduction of said vector into a host cell is higher than when only one of said intron-originated DNA fragments is inserted into a site upstream of said foreign gene.

Details for Patent 6,214,578

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		2016-06-12
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	For Injection	103132			2016-06-12
Merck Sharp & Dohme Corp.	INTRON A	interferon alfa-2b	Injection	103132			2016-06-12