A COMMON MULTIPLE CLONING SITE IN A SET OF VECTORS FOR EXPRESSION OF EUKARYOTIC GENES IN MAMMALIAN, INSECT AND BACTERIAL-CELLS

Here, we describe the construction of plasmid vectors facilitating exp ression of cloned genes in bacteria and in cells of mammalian and inse ct origin. Two types of multiple cloning site (MCS) were designed base d on the MCS in the expression vector lambda gt11Sfi-Not. In the first set of vectors a start Met codon was included in the same reading fra me as in lambda t11Sfi-Not to support expression of partial cDNA clone s. Thus a cDNA insert of lambda t11Sfi-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 Me t codon. By Western blot analysis and by transactivation of a reporter plasmid in co-transfections we show that cDNA is very efficiently exp ressed in NIH 3T3 cells under control of the elongation factor la prom oter.