title: Alkaline phosphatase inhibitors as labels of DNA probes.
abstract: A new approach to nucleic acid labeling was developed by preparing bifunctional reagents containing, in addition to the DNA-linking group, a competitive inhibitor of the chromogenic enzyme alkaline phosphatase. The nucleic acids labeled in such a way were able to bind themselves to the enzyme, whose activity was restored in the presence of a chromogenic substrate. Five phosphonic-acid-containing reagents were synthesized and coupled to linearized pBR322 plasmid DNA by different condensation methods. Eight probes thus obtained were assayed in a modified dot-blot detection procedure obtaining the best nucleic acid detection sensitivity of 25 pg. Finally, five of the above probes were tested in hybridization experiments, reaching sensitivity of 50 pg.
authors.forename: E
authors.initials: E
authors.lastname: Davini
authors.forename: C
authors.initials: C
authors.lastname: Di Leo
authors.forename: A
authors.initials: A
authors.lastname: Rossodivita
authors.forename: P
authors.initials: P
authors.lastname: Zappelli
mesh_terms: D000469:Alkaline Phosphatase / Q000037:antagonists & inhibitors*; D002863:Chromogenic Compounds / Q000737:chemistry; D015342:DNA Probes / Q000737:chemistry*; D004791:Enzyme Inhibitors / Q000737:chemistry*; D007700:Kinetics; D009693:Nucleic Acid Hybridization; D063065:Organophosphonates / Q000737:chemistry*; D010957:Plasmids / Q000235:genetics; D012680:Sensitivity and Specificity
chemical_list: D002863:Chromogenic Compounds; D015342:DNA Probes; D004791:Enzyme Inhibitors; D063065:Organophosphonates; D000469:Alkaline Phosphatase