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