New nucleoside tri:phosphoric acid ester derivs. - used for prepn. of polymerase chain reaction probes, amplification of nucleic acid sequences and as chain terminators for sequences

摘要

New labelled nucleoside triphosphate derivs. (I) are ribonucleosides, 2'-deoxyribonucleosides or 2',4'-dideoxyribonucleosides, opt. in salt form, with a gp. of formula -(A)nQ linked to the nucleoside base, where: n = 0 or 1; A = an optional linking gp. stated to be an amino, alkyl, alkenyl, alkynyl, aryl, aralkyl or alkaryl gp. which may contain one or more terminal or internal structural functional units selected from amino, amido, imino, hydrazino, azo, triazene, keto, ester, hydroxy, ether, thioether, disulphide, ureido and thioureido gps., opt. substd. by alkyl, alkenyl, aryl, alkaryl or aralkyl gps.; Q = a gp. which is capable of complexing lanthanide ions and contains an 'energy donor' for lanthanide ions. Lanthanide complexes of (I) are also new. USE/ADVANTAGE - The complexes may be incorporated in oligonucleotides, e.g. by enzyme-catalysed tailing (using terminal deoxynucleotidyl transferase), to produce fluorescence-labelled hydridisation probes. They may also be used as markers in PCR amplification systems, as chain terminators (in the case of the dideoxy derivs.) in nucleic acid sequencing systems, and as tracers for measuring nucleic acid synthesis in vitro or in vivo. The lanthanide label can be detected by time-resolved fluorometry, which eliminates interference from autofluorescence and light scattering.