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Last Updated: April 24, 2024

Claims for Patent: 5,770,400


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Summary for Patent: 5,770,400
Title: Exogenous gene expression vector containing chick .beta.-actin gene promoter
Abstract:An expression vector for the expression of an exogenous gene in an animal cell, which contains a chick .beta.-actin gene promoter and a restriction enzyme site for incorporating an exogenous gene at the downstream of said promoter and a process for expressing an exogenous gene, which comprises incorporating said exogenous gene into the expression vector at the restriction enzyme site for incorporating the exogenous gene, introducing said vector into an animal cell and culturing the obtained transformed animal cell. The expression system of the present invention can be applied to an expression of any exogenous gene and has an extremely high expression efficiency and is applicable to a wide range of host cells, and hence can sufficiently be utilized for industrial-scale production of a useful material.
Inventor(s): Miyazaki; Junichi (Kumamoto, JP), Yamamura; Ken-ichi (Kumamoto, JP), Araki; Masatake (Kumamoto, JP), Yonemura; Hiroshi (Kumamoto, JP), Nozaki; Chikateru (Kumamoto, JP)
Assignee: Juridical Foundation The Chemo-Sero-Therapeutic Research Institute (Kumamoto, JP)
Application Number:08/814,468
Patent Claims:1. An expression vector for the expression of an exogenous gene in an animal cell, wherein said vector comprises a modified chick .beta.-actin gene promoter and a restriction enzyme site for incorporating an exogenous gene downstream of said chick .beta.-actin gene promoter, wherein said modified chick .beta.-actin promoter is a hybrid promoter formed by:

deleting a DNA fragment from the 3' end of said promoter, wherein said fragment (A) ranges from the middle of the intron of said promoter to a location downstream thereof and (B) comprises a splicing acceptor sequence, and

linking to the 3' end of said promoter a DNA fragment containing an alternate splicing acceptor sequence.

2. The expression vector of claim 1, wherein said alternate splicing acceptor sequence is a DNA fragment containing a splicing acceptor sequence from a rabbit .beta.-globin gene.

3. The expression vector of claim 1, wherein said expression vector further comprises at least one gene derived from E. coli for cloning said vector in E. coli.

4. The expression vector of claim 3, wherein said at least one gene is a drug resistant gene and/or a replication origin capable of functioning in E. coli.

5. The expression vector of claim 1, wherein said expression vector further comprises an SV 40 early transcription splicing sequence and a polyadenylation sequence.

6. The expression vector of claim 5, wherein said expression vector further comprises the SV 40 replication origin.

7. The expression vector of claim 1, wherein said expression vector further comprises the SV 40 replication origin.

8. A process for expressing an exogenous gene which comprises

(a) incorporating said exogenous gene into an expression vector for the expression of an exogenous gene in an animal cell, the expression vector comprising a modified chick .beta.-actin gene promoter and a restriction enzyme site for incorporating an exogenous gene downstream of said chick .beta.-actin gene promoter,

(b) introducing said vector into an animal cell and

(c) culturing the resulting transformed cell, wherein said modified chick .beta.-actin promoter is a hybrid promoter formed by:

deleting a DNA fragment from the 3' end of said promoter, wherein said fragment (A) ranges from the middle of the intron of said promoter to a location downstream thereof and (B) comprises a splicing acceptor sequence, and

linking to the 3' end of said promoter a DNA fragment containing an alternate splicing acceptor sequence.

9. The process of claim 8, wherein said expression vector further comprises at least one gene derived from E. coli for cloning said vector in E. coli.

10. The process of claim 8, wherein said expression vector further comprises an SV 40 early transcription splicing sequence and polyadenylation sequence.

11. The process of claim 8, wherein said expression vector further comprises the SV 40 replication origin.

Details for Patent 5,770,400

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 2008-06-24
Merck Sharp & Dohme Corp. INTRON A interferon alfa-2b For Injection 103132 ⤷  Try a Trial 2008-06-24
Merck Sharp & Dohme Corp. INTRON A interferon alfa-2b Injection 103132 ⤷  Try a Trial 2008-06-24
>Applicant >Tradename >Biologic Ingredient >Dosage Form >BLA >Approval Date >Patent No. >Expiredate

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