Method of using eukaryotic expression vectors comprising a poly

Details Method of using eukaryotic expression vectors comprising a poly Method of using eukaryotic expression vectors comprising a poly

1993-08-23

1996-04-09

Fleisher, Mindy

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Recombinant dna technique included in method of making a...

4352402, 4353201, C12P 2102, C12N 510, C12N 1585

Patent

active

055061184

ABSTRACT:
The present invention is a method of using a poly-GT element with a eukaryotic promoter in the presence of an immediate-early gene product of a large DNA virus to increase 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. A novel enhancer system is described comprising a cis-acting poly-GT element and a trans-acting E1A gene product, whereby the poly-GT element does not itself possess enhancer activity with certain eukaryotic promoters but rather requires the E1A gene product for enhancer activity. The present invention further comprises a number of useful expression vectors that comprise a poly-GT element with a 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, tissue plasminogen activator, a modified tissue plasminogen activator, or an interferon. The present invention also comprises a method for further increasing the expression of a useful substance involving a poly-GT element, the E1A gene product, and a BK enhancer which has been placed immediately upstream (within 0 to about 300 nucleotides) of the eukaryotic promoter used in tandem with a BK enhancer.

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