High-copy expression vector based on amplification-promoting sequences.

Abstract:

:We describe a new vector system that allows efficient expression of heterologous proteins in transformed mouse L fibroblasts. This is due to its persistence at high copy numbers, achieved by a 370-bp amplification promoting element (muNTS1) derived from the nontranscribed spacer of murine rDNA. Copy number determination showed that this sequence mediates a 40- to 800-fold amplification of the vector DNA in transfected L cells. High copy number was accompanied by increased expression levels of the reporter gene secreted alkaline phosphatase (SEAP). Analyzing the structural organization of multicopy plasmid DNA in mouse L cells revealed that plasmid DNA is integrated as reiterated head-to-tail concatamers into the chromosomal DNA. The vector described here can be used as a versatile high-copy expression system for heterologous proteins overcoming any limitation to enzyme-deficient cell lines.

journal_name

DNA Cell Biol

journal_title

DNA and cell biology

authors

Hemann C,Gärtner E,Weidle UH,Grummt F

doi

10.1089/dna.1994.13.437

subject

Has Abstract

pub_date

1994-04-01 00:00:00

pages

437-45

issue

4

eissn

1044-5498

issn

1557-7430

journal_volume

13

pub_type

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