NEW VECTORS FOR THE IN-VITRO GENERATION OF ALKALINE-PHOSPHATASE FUSIONS TO PROTEINS ENCODED BY G-RICH DNA(C)

Phagemid vectors were constructed to allow fusions of alkaline phospha tase to proteins encoded by G + C-rich DNA, by engineering a BstBI sit e (TT/CGAA) in front of a phoA gene that lacks an encoded signal pepti de. Three vectors (pJDT1, pJDT2 and pJDT3), each with phoA in a differ ent reading frame with respect to the BstBI site, were produced; a lac P region is present in each plasmid upstream of the BstBI site. The pr esence of the BstBI site allows the random cloning of G + C-rich DNA d igested with a number of restriction enzymes that generate cohesive en ds.