HSV helper virus independent vector

Abstract

This invention encompasses an eukaryotic expression vector constructed from a prokaryotic plasmid by inserting into the prokaryotic plasmid a mediator sequence and a promoter/regulatory sequence with a restriction endonuclease site for inserting a gene to be expressed. The gene to be expressed is inserted into the restriction endonuclease site in the promoter/regulatory sequence and the resulting plasmid is in turn used to transform a eukaryotic cell. The gene is expressed with or without viral mediation.

Description

Claims

1. A process for producing a mammalian cell line having cells containing extrachromosomal eukaryotic expression vectors, which process comprises the steps of:

(a) inserting a herpes simplex virus (HSV) BamHI N fragment into a prokaryotic plasmid in a manner which does not disrupt the prokaryotic plasmid function to form a BamHI N fragment-containing plasmid;

(b) adding a eukaryotic promoter and a gene to be expressed to the product of (a), said promoter and said gene to be expressed being operably linked to each other, and said promoter being located immediately 5' of said gene to be expressed;

(c) introducing the plasmid of (b) into a mammalian cell; and

(d) propagating said mammalian cell in the absence of HSV.

2. A process according to claim 1, wherein said mammalian cell is a baby hamster kidney cell.

3. A process according to claim 1, wherein said mammalian cell is a mouse Ltk- cell.

4. The process of claim 1 wherein said mammalian cell is propagated for at least thirty cell generations in he absence of HSV.