Synthesis of fluorophore-labeled DNA

Details Synthesis of fluorophore-labeled DNA Synthesis of fluorophore-labeled DNA

1997-11-07

1999-08-31

Knode, Marian C.

Chemistry: molecular biology and microbiology

Micro-organism, tissue cell culture or enzyme using process...

Preparing compound containing saccharide radical

435 6, 435270, 536 231, 536 243, 436 94, 436173, C12P 1934

Patent

active

059453125

ABSTRACT:
Sequencing methods and methods for synthesizing DNA probes using mutant bacteriophage T4 DNA polymerases which have increased ability to incorporate modified nucleotides for the synthesis of long or short chains of complementary, modified, e.g., fluorophore-labeled DNA. In general, the mutant T4 DNA polymerases retain 3'.fwdarw.5' exonuclease activity; hence, reduction or elimination of 3'.fwdarw.5' exonuclease activity is not a prerequisite for efficient synthesis of a complementary fluorophore-labeled or other modified DNA. In fact, retention of 3'.fwdarw.5' exonuclease activity increases accuracy of DNA replication, because these exonucleases proofread or edit the product of DNA replication.

REFERENCES:
patent: 4711955 (1987-12-01), Ward et al.
Delarue et al., "An Attempt to Unify the Structure of Polymerase," Protein Engineering, 3(6), 461-467 (1990).
Reha-Krantz et al.(I), "Bacteriophage T4 DNA Polymerase Mutations That Confer Sensitivity to the PP.sup.i Analog Phosphonoacetic Acid," J. Virology, 67(1), 60-66 (Jan. 1993).
Reha-Krantz et al.(II), "Genetic and Biochemical Studies of Bacteriophage T4 DNA Polymerase 3'.fwdarw.5'-Exonuclease Activity," J. Biological Chemistry, 268(36), 27100-27108 (Dec. 1993).
Stocki et al., "Dynamics of Bacteriophage T4 DNA Polymerase Function: Identification of Amino Acid Residues that Affect Switching Between Polymerase and 3'.fwdarw.5' Exonuclease Activity," J. Molecular Biology, 254, 15-28 (1995).
Delarue, M., et al., "An attempt to unify the structure of polymerases", Protein Engineering, 3(6)461-467 (1990).
Reha-Krantz, L. J., et al., "Bacteriophage T4 DNA Polymerase Mutations That Confer Sensitivity to the PP.sup.i Analog Phosphonoacetic Acid", J. Virol., 67(1):60-66 (1993) (Jan.).
Reha-Krantz, L. J., et al., "Genetic and Biochemical Studies of Pacteriophage T4 DNA Polymerase 3'.fwdarw.5'-Exonuclease Activity", J. Biol. Chem., 268(36):27100-27108 (1993)(Dec.).
Stocki, S.A., et al., "Dynamics of Bacteriophage T4 DNA Polymerase Function: Identication of Amino Acid Residues that Affect Switching between Polymerase and 3'.fwdarw.5' Exonuclease Activites", J. Mol. Biol., 254:15-28 (1995).

Affiliated with

Also associated with

Rating

Synthesis of fluorophore-labeled DNA does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Synthesis of fluorophore-labeled DNA, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Synthesis of fluorophore-labeled DNA will most certainly appreciate the feedback.

Rate now

Comments { 0 }

Profile ID: LFUS-PAI-O-2426360