US 5,858,704 A · Method of using eukaryotic expression vectors comprising the BK virus enhancer

Abstract

The present invention is a method of using the BK enhancer in tandem with a eukaryotic promoter to promote transcription of DNA that encodes a useful substance. The method of the present invention requires the presence of the E1A gene product for maximum expression of the useful substance. The present invention also comprises a number of useful expression vectors that comprise the BK enhancer in tandem with the adenovirus 2 late promoter positioned to drive expression of a variety of proteins, such as protein C, chloramphenicol acetyltransferase, and tissue plasminogen activator. The present invention further comprises a method for increasing the activity of the BK enhancer involving placement of the BK enhancer immediately upstream of the eukaryotic promoter used in tandem with the BK enhancer to drive expression of a useful substance. Furthermore, the present invention also comprises a method for coamplification of genes in primate cells. Additionally, the invention further comprises the recombinant human protein C molecule produced in 293 cells which comprises novel glycosylation patterns.

Claims4

Dependent claims

In a method for producing a protein that is naturally .gamma.-carboxylated, properly folded and processed and wherein said protein is encoded in a recombinant DNA vector such that said protein is expressed when a eukaryotic host cell containing said vector is cultured under suitable expression conditions, the improvement comprising:

(a) inserting said vector into an adenovirus-transformed host cell;
and
(b) culturing said host cell of step a) under growth conditions and in media containing sufficient vitamin K for carboxylation.

Dependencies are inferred from the claim text.

Facts

Application

08/459,644Filed 2 Jun 1995 · division

Priority

9 Apr 1986

Granted

12 Jan 1999

Term ends

Not estimatedThe source records don’t support an estimate.

Classification18 codes

C12N 9/6464
Serine endopeptidases: protein C
C12N 1/38
Microorganisms; Compositions thereof ; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of…:…
C12N 9/1033
Transferring groups other than amino-acyl groups: chloramphenicol O-acetyltransferase
C12N 9/6459
Plasminogen activators: t-plasminogen activator , i.e. tPA

Family

Serial849,999Parent of this continuation-in-part · filed 9 Apr 1986

Serial129,028Parent of this continuation-in-part · filed 4 Dec 1987

Serial250,001Parent of this continuation-in-part · filed 27 Sep 1988

Serial368,700Parent of this continuation · filed 20 Jun 1989

Serial208,930Parent of this division · filed 9 Mar 1994

US 5,550,036 AParent of this division

US 5,858,704 AThis patent

From the US filing history. Foreign family members aren’t listed.

Method of using eukaryotic expression vectors comprising the BK virus enhancer

Life of this patent

Method of using eukaryotic expression vectors comprising the BK virus enhancer

Life of this patent

Abstract

Claims4