Bicistronic Vector for the Creation of Stable Mammalian Cell Lines that Predisposes All Antibiotic-Resistant Cells to Express Recombinant Protein. (January 1996)

• Record Type: Journal Article
• Authors: Rees, S.; Coote, J.; Stables, J.; Goodson, S.; Harris, S.; Lee, M.G.
• Is Part Of: Biotechniques. Volume 20:Number 1(1996)
• Page Start: 102
• Page End: 110
• Publication Date: 1996-01
• Subjects: Biology, Experimental -- Periodicals; Molecular biology -- Periodicals; Medical technology -- Periodicals; Biology, Experimental; Medical technology; Molecular biology; Clinical Laboratory Techniques -- Periodicals; Research -- Periodicals; Medical Laboratory Science -- Periodicals; Periodicals; Electronic journals; 570
• DOI: 10.2144/96201st05
• Languages: English
• ISSNs: 0736-6205

Abstract

We have developed an improved vector for the stable expression of recombinant protein in mammalian cells. In this vector, designated pCIN, both the recombinant cDNA and the neomycin phosphotransferase selection marker are transcribed from a single promoter element. To facilitate translation of the second open reading frame, the encephalomyocarditis virus internal ribosome entry site has been inserted into the expression cassette immediately before the start codon of this sequence. We report the use of this vector to generate stable cell lines expressing the human 5-HT1D α serotonin receptor and show that following transfection and clona selection, all ten cell lines characterized express similar and high levels of receptor (1.5–11.9 pmol receptor/mg protein). Use of pCIN should permit the rapid and efficient production of stable mammalian cell lines for the characterization of recombinant protein, as this vector appears to predispose all transfected cells to express such protein.