title: New vectors for the in vitro generation of alkaline phosphatase fusions to proteins encoded by G+C-rich DNA.
abstract: Phagemid vectors were constructed to allow fusions of alkaline phosphatase to proteins encoded by G+C-rich DNA, by engineering a BstBI site (TT/CGAA) in front of a phoA gene that lacks an encoded signal peptide. Three vectors (pJDT1, pJDT2 and pJDT3), each with phoA in a different reading frame with respect to the BstBI site, were produced; a lacP region is present in each plasmid upstream of the BstBI site. The presence of the BstBI site allows the random cloning of G+C-rich DNA digested with a number of restriction enzymes that generate cohesive ends.
authors.affiliation: Department of Biochemistry and Microbiology, University of Victoria, British Columbia, Canada.
authors.forename: K E
authors.initials: KE
authors.lastname: Mdluli
authors.forename: J D
authors.initials: JD
authors.lastname: Treit
authors.forename: V J
authors.initials: VJ
authors.lastname: Kerr
authors.forename: F E
authors.initials: FE
authors.lastname: Nano
mesh_terms: D000469:Alkaline Phosphatase / Q000096:biosynthesis* / Q000235:genetics; D000595:Amino Acid Sequence; D001483:Base Sequence; D004269:DNA, Bacterial / Q000235:genetics; D005822:Genetic Vectors; D008969:Molecular Sequence Data; D009169:Mycobacterium tuberculosis / Q000235:genetics; D016133:Polymerase Chain Reaction; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D004269:DNA, Bacterial; D011993:Recombinant Fusion Proteins; D000469:Alkaline Phosphatase