TY - JOUR
T1 - [25] Purification and Properties of a Pyrimidine Dimer-Specific Endonuclease from E. Coli Infected with Bacteriophage T4
AU - Friedberg, Errol C.
AU - Ganesan, Ann K.
AU - Seawell, Patricia C.
N1 - Funding Information:
This work was supported by grants from the United States Public Health Service (CA 12428 and GM 19010), the American Cancer Society (NPI61B and NP 174), and by con-tracts with the United States Department of Energy \[EY-76-5-03-0326 (PA7 and 32)\]. We thank Richard Cone for instruction in several of the analytical procedures. K. Sato and M. Sekiguchi, J. Mol. Biol. 102, 15 (1976). a S. Yasuda and M. Sekiguchi, Proc. Natl. Acad. Sci. U.S.A. 67, 1839 (1970).
Funding Information:
Investigations were supported by research grants from the American Cancer Society (NPI26) and the National Cancer Institute (CA16519) and by a training grant from the latter (CA90139). 2 C. C. Richardson, I. R. Lehman, and A. Kornberg, J. Biol. Chem. 239, 251 (1964). a C. C. Richardson and A. Kornbcrg, J. Biol. Chem. 239, 242 (1964). 4 This activity has also been called cndonucleasc VI and is one of at least two different activitiest hat have been referred to as endonuclease II. The preferred designation is "AP endonuclease activity of cxonuclease HI." 5 B. Weiss, S. G. Rogers, and A. F. Taylor, in "DNA Repair Mechanisms: ICN-UCLA Symposia on Molecular and Cellular Biology" (P. C. Hanawalt, E. C. Fdedbcrg, and C. F. Fox, eds.), p. 191. Academic Press, New York, 1978.
PY - 1980/1/1
Y1 - 1980/1/1
N2 - The chapter explains purification and properties of a pyrimidine dimer-specific endonuclease from E. coli infected with bacteriophage T4. The pyrimidine dimer-specific endonuclease of bacteriophage T4 is coded by the v (denV) gene. The enzyme has been called endonuclease V of phage T4 or the T4 uv endonuclease. For detecting pyrimidine dimers a preparation of enzyme free of activity against unirradiated DNA or DNA containing other types of damage is required. The chapter describes several methods for assaying T4 endonuclease V as well as the partial purification and general properties of the enzyme. The chapter discusses a reproducible procedure for obtaining stable preparations of the enzyme which can be used as a specific probe for pyrimidine dimers. For the purpose of purification, the enzyme can be assayed by one of three general methods— namely, coupled nuclease assay, filter binding assay, and DNA nicking assay. The chapter further discusses enzyme purification.
AB - The chapter explains purification and properties of a pyrimidine dimer-specific endonuclease from E. coli infected with bacteriophage T4. The pyrimidine dimer-specific endonuclease of bacteriophage T4 is coded by the v (denV) gene. The enzyme has been called endonuclease V of phage T4 or the T4 uv endonuclease. For detecting pyrimidine dimers a preparation of enzyme free of activity against unirradiated DNA or DNA containing other types of damage is required. The chapter describes several methods for assaying T4 endonuclease V as well as the partial purification and general properties of the enzyme. The chapter discusses a reproducible procedure for obtaining stable preparations of the enzyme which can be used as a specific probe for pyrimidine dimers. For the purpose of purification, the enzyme can be assayed by one of three general methods— namely, coupled nuclease assay, filter binding assay, and DNA nicking assay. The chapter further discusses enzyme purification.
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U2 - 10.1016/S0076-6879(80)65027-7
DO - 10.1016/S0076-6879(80)65027-7
M3 - Article
C2 - 6246342
AN - SCOPUS:0018848399
VL - 65
SP - 191
EP - 201
JO - Methods in Enzymology
JF - Methods in Enzymology
SN - 0076-6879
IS - C
ER -