New vectors for the in vitro generation of alkaline phosphatase fusions to proteins encoded by G+C-rich DNA.
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.
Mdluli
K E
KE
Department of Biochemistry and Microbiology, University of Victoria, British Columbia, Canada.
Treit
J D
JD
Kerr
V J
VJ
Nano
F E
FE
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
D004269:DNA, Bacterial; D011993:Recombinant Fusion Proteins; D000469:Alkaline Phosphatase