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Yeung Anthony T
Polynucleotides encoding mismatch endonuclease CELI and diagnostic and therapeutic uses thereof
. 2001 20010222. :93 pp
PMID: AN 2001:636264   
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Abstract
The invention claims polynucleotide and polypeptide sequences for a mismatch endonuclease named CEL I and a purified, active 39 kD protein which is a putative CEL II isoenzyme. Use of CEL I polynucleotide sequences in recombinant prodn. of an endonuclease, genetic vectors, and host cells are claimed. In addn., methods for screening compds. which modulate CEL I endonuclease activity and antibodies specific for an endonuclease are claimed. The invention further claims methods for use of CEL I endonuclease for the detection of mutations in targeted polynucleotide sequences, to facilitate the localization and identification of mutations, mismatches and genetic polymorphisms. Methods for detecting mutations using CEL I endonuclease involve exposing hybridized, labeled polynucleotides to the enzyme and identifying the cleaved enzyme products in the cases where mismatched sequences were present. Amino acid sequences encoding Arabidopsis thaliana BFN1 (bifunctional nuclease 1), Zinnia elegans endonuclease ZEN1, and Hemerocallis DSA6 (daylily senescence-assocd. protein 6) are greater than 60% identical to CEL I endonuclease and use of these polypeptides by the methods of this invention is claimed. Mutations in the BRCA1 gene were analyzed using CEL I endonuclease and Genescan fragment anal. on an automated DNA sequencer. BRCA1 DNA was amplified using PCR and one strand was labeled with 5'-FAM, the other with TET. Two-color heterduplexes were incubated with CEL I mismatch endonuclease and mismatch-specific cleavage was obsd. [on SciFinder (R)]
Notes
CAN 135:206492 3-3 Biochemical Genetics Patent written in English. 20010830 52906-91-9P (nuclease CEL I) Role: ARG (Analytical reagent use), BAC (Biological activity or effector, except adverse), BPN (Biosynthetic preparation), BSU (Biological study, unclassified), PRP (Properties), PUR (Purification or recovery), THU (Therapeutic use), ANST (Analytical study), BIOL (Biological study), PREP (Preparation), USES (Uses) (CEL I isoenzyme; polynucleotides encoding mismatch endonuclease CEL1 and diagnostic and therapeutic uses thereof); 358016-03-2P Role: ARG (Analytical reagent use), BAC (Biological activity or effector, except adverse), BPN (Biosynthetic preparation), BSU (Biological study, unclassified), PRP (Properties), PUR (Purification or recovery), THU (Therapeutic use), ANST (Analytical study), BIOL (Biological study), PREP (Preparation), USES (Uses) (amino acid sequence; polynucleotides encoding mismatch endonuclease CEL1 and diagnostic and therapeutic uses thereof); 358016-02-1 Role: BUU (Biological use, unclassified), PRP (Properties), BIOL (Biological study), USES (Uses) (nucleotide sequence; polynucleotides encoding mismatch endonuclease CEL1 and diagnostic and therapeutic uses thereof); 358018-02-7; 358018-03-8; 358018-04-9; 358018-05-0; 358018-06-1; 358350-33-1; 358350-35-3 Role: PRP (Properties) (unclaimed protein sequence; polynucleotides encoding mismatch endonuclease CELI and diagnostic and therapeutic uses thereof); 357915-90-3; 357915-91-4 Role: PRP (Properties) (unclaimed sequence; polynucleotides encoding mismatch endonuclease CELI and diagnostic and therapeutic uses thereof) A1 US 2000-510322 20000222