Preparation of oligodeoxynucleotide-alkaline phosphatase conjugates and their use hybridization probes

摘要: Abstract Short synthetic oligonucleotides have been covalently cross-linked to alkaline phosphatase using the homobifunctional reagent disuccinimidyl suberate. The oligomers, twenty-one twenty-six bases in length, are complementary unique sequences found herpes simplex virus, hepatitis B Campylobacter jejuni and enterotoxigenic Escherichia coli. Each oligomer contains a single modified base with 12-atom "linker arm" terminating reactive primary amine. Cross-linking through this amine results oligomer-enzyme conjugates composed of one per enzyme molecule that full activity can hybridize target DNA fixed nitrocellulose within 15 minutes. hybrids detected directly dye precipitation assay at sensitivity 10(6) molecules (2 X 10(-18) mol) 4 hours development time. has no apparent effect on selectivity or kinetics oligonucleotide hybridization be hybridized melted off conventional manner.