NEW LAMBDA AND PLASMID VECTORS FOR EXPRESSION CLONING IN MAMMALIAN-CELLS

This report describes the construction of a new family of lambda phage and plasmid cloning vectors. lambda GDST3/T7 allows cDNA insertion li p to 14 kb; it is derived from lambda NM1151 by the insertion of a mul tiple cloning site containing eight unique restriction sires. The two asymmetrical SfiI sites are flanked by the T3 and T7 promoters direct sequencing and in vitro transcription/translation. The same multiple c loning site is also present in both orientations in the eukaryotic exp ression plasmids, pGDSV3 and pGDSV7. By exploiting the superior discri mination of the signal-to-noise ratio of the lambda vectors for primar y screening (by either nucleic acids or antibody probes), relevant cDN As can thus be efficiently transferred through SfiI sites into the pla smids pGDSV3/7 for functional secondary screening by expression in mam malian cells.