Methods and compositions for optical detection of...

Chemistry: molecular biology and microbiology – Measuring or testing process involving enzymes or... – Involving nucleic acid

Reexamination Certificate

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C530S350000

Reexamination Certificate

active

07544469

ABSTRACT:
New Gp32F protein for determining, localizing or quantitating single-stranded DNA or RNA molecules comprises a Gp32 protein conjugated at an environment-sensitive amino acid (e.g. cysteine-166) to a fluorescent label.

REFERENCES:
Villemain et al. 1993; Energetics of arginine-4 substitution mutants in the N-terminal cooperativity domain of T4 gene 32 protein. Biochemistry 32: 11235-11246.
Xu et al. 2001; Helicase assembly protein Gp59 of bacteriophage T4: Fluorescence anisotropy and sedimentation studies of complexes formed with derivatives of Gp32, the phage ssDNA binding protein. Biochemistry 40: 7651-7661.
en.wikipedia.org/wiki/Fluorescent—labelling; Dec. 12, 2008.
Chase, J.W. et al. Single-stranded DNA binding proteins required for DNA replication. Annu Rev Biochem. 55: 103-136, 1986.
Harris, L.D. et al. Formation of D loops by the UvsX protein of T4 bacteriophage: a comparison of the reaction catalyzed in the presence of absence of gene 32 protein. Biochemistry. 27(18): 6954-6959, 1988.
Huang, H. et al. Fidelity and predominant mutations produced by deep vent wild-type and exonuclease-deficient DNA polymerases during in vitro DNA amplification. DNA Cell Biol. 15(7): 589-594, 1996.
Kelly et al. DNA “melting” proteins III: Flourescence “mapping” of the nucleic acid binding site of bacteriophage T4 gene 32-protein. Journal of Biological Chemistry 251(22): 7229-7239, 1976.
Kelly et al. DNA “melting” proteins IV: Flourescence measurements of binding parameters for bacteriophage T4 gene 32-protein to mono-, oilgo-, and polynucleotides. Journal of Biological Chemistry 251(22): 7240-7250, 1976.
Kowalczykowski, S.C. et al. Cooperative and noncooperative binding of protein ligands to nucleic acid lattices: experimental approaches to the determination of thermodynamic parameters. Biochemistry. 25(6): 1226-1240, 1986.
Kroutil, L.C. et al. Effect of accessory proteins on T4 DNA polymerase replication fidelity. J Mol Biol. 278(1): 135-146, 1998.
Muniyappa, K. et al. Mechanism of the concerted action of recA protein and helix-destabilizing proteins in homologous recombination. Proc Natl Acad Sci U S A. 81(9): 2757-2761, 1984.
Shamoo, Y., et al. Crystal structure of a replication fork single-stranded DNA binding protein (T4 gp32) complexed to DNA. Nature. 376(6538): 362-366, 1995.
Villemain, J.L. et al. Mutations in the N-terminal cooperativity domain of gene 32 protein alter properties of the T4 DNA replication and recombination systems, J Biol Chem. 275(40): 31496-31504, 2000.
Yonesaki, T. et al. Synergistic action of three recombination gene products of bacteriophage T4, uvsX, uvsY, and gene 32 proteins. J Biol Chem. 264(14): 7814-7820, 1989.

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