A common multiple cloning site in a set of vectors for expression of eukaryotic genes in mammalian, insect and bacterial cells

Niels Pallisgaard, Finn Skou Pedersen, Svend Birkelund, Poul Jørgensen*

*Corresponding author

Publikation: Bidrag til tidsskriftTidsskriftartikelpeer review

Abstract

Here, we describe the construction of plasmid vectors facilitating expression of cloned genes in bacteria and in cells of mammalian and insect origin. Two types of multiple cloning site (MCS) were designed based on the MCS in the expression vector λgt11Sfi-Not. In the first set of vectors a start Met codon was included in the same reading frame as in λgt11Sfi-Not to support expression of partial cDNA clones. Thus a cDNA insert of λgt11Sfi-Not could be shuttled among the new vectors for expression. The other set of vectors without a start codon were suitable for expression of cDNA carrying their own start Met codon. By Western blot analysis and by transactivation of a reporter plasmid in co-transfections we show that cDNA is very efficiently expressed in NIH 3T3 cells under control of the elongation factor 1α promoter.

OriginalsprogEngelsk
TidsskriftGene
Vol/bind138
Udgave nummer1-2
Sider (fra-til)115-118
Antal sider4
ISSN0378-1119
DOI
StatusUdgivet - 28 jan. 1994
Udgivet eksterntJa

Emneord

  • Antibody detection
  • bacteriophage λ
  • E. coli, eukaryotic and prokaryotic vectors
  • transactivation
  • transcription factors

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