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Exonucleases

Larry A. McReynolds¹, Nicole M. Nichols¹

¹New England Biolabs, Ipswich, Massachusetts

Publication Name:

Current Protocols in Molecular Biology

Unit Number:

Unit 3.11

DOI:

10.1002/0471142727.mb0311s96

Online Posting Date:

October, 2011

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Abstract

Reaction conditions for numerous exonucleases are detailed in this unit along with discussions of potential applications and preferred substrates. Single-stranded and double-stranded 5¢ 3¢ and 3¢ 5¢ exonucleases are included. Curr. Protoc. Mol. Biol. 96:3.11.1-3.11.7. © 2011 by John Wiley & Sons, Inc.

Keywords: DNases; ssDNA exonucleases; dsDNA exonucleases

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Introduction
Single-Stranded 3¢ 5¢ Exonucleases
Single-Stranded 5¢ 3¢ and 3¢ 5¢ Exonucleases
Double-Stranded 5¢ 3¢ Exonucleases
Double-Stranded 3¢ 5¢ Exonuclease
Single- and Double-Stranded 5¢ 3¢ and 3¢ 5¢ Exonuclease
Literature Cited
Figures
Tables

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Figures

Figure 3.11.1

Exonuclease VII activity.

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Figure 3.11.2

5¢ 3¢ exonuclease activity: T7 gene 6 exonuclease and exonuclease.

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Figure 3.11.3

Exonuclease III activity.

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Literature Cited

Literature Cited
	Berk, A.J. and Sharp, P.A. 1977. Sizing and mapping of early adenovirus mRNAs by gel electrophoresis of S1 endonuclease digested hybrids. Cell 12:721-732.
	Demple, B., Johnson, A., and Fung, D. 1986. Exonuclease III and endonuclease IV remove 3¢ blocks from DNA synthesis primers in H₂O₂-damaged Escherichia coli. Proc. Natl. Acad. Sci. U.S.A. 83:7731-7735.
	Eichler, D.C. and Lehman, I.R. 1977. On the role of ATP in phosphodiester bond hydrolysis catalyzed by the recBC deoxyribonuclease of Esherichia coli. J. Biol. Chem. 252:499-503.
	Goff, S. and Berg, P. 1978. Excision of DNA segments introduced into cloning vectors by the poly(dA-dT) joining method. Proc. Natl. Acad. Sci. U.S.A. 75:1763-1767.
	Henikoff, S. 1984. Unidirectional digestion with exonuclease III creates targeted breakpoints for DNA sequencing. Gene 28:351-359.
	Karu, A., MacKay, V., Goldmark, P., and Linn, S. 1973. The recBC deoxyribonuclease of Escherichia coli K-12. Substrate specificity and reaction intermediates. J. Biol. Chem. 248:4874-4884.
	Kerr, C. and Sadowski, P.D. 1972. Gene 6 exonuclease of bacteriophage T7. I. Purification and properties of the enzyme. J. Biol. Chem. 247:305-310.
	Krishnakumar, S., Zheng, J., Wilhelmy, J., Faham, M., Mindrinos, M., and Davis, R. 2008. A comprehensive assay for targeted multiplex amplification of human DNA sequences. Proc. Natl. Acad Sci. U.S.A. 105:9296-9301.
	Lehman, I.R. and Nussbaum, A.L. 1964. The deoxyribonucleases of Escherichia coli V. on the specificity of exonuclease I (phosphodiesterase). J. Biol. Chem. 239:26-28.
	Li, H., Cui, X. and Arnheim, N. 1991. Eliminating primers from completed polymerase chain reactions with exonuclease VII. Nucleic Acids Res. 19:3139-3141.
	Little, J.W. 1981. "Lambda exonuclease". In Gene Amplification and Analysis, Vol. 2: Structural analysis of nucleic acids ( J.G. Chirikjian and T.S. Papas, eds.) pp. 135-145. Elsevier/North Holland, NY, Amsterdam, Oxford.
	Rogers, S.G. and Weiss, B. 1980. Exonuclease III of Escherichia coli K-12, an AP exonuclease. Methods Enzymol. 65:201-211.
	Sanger, F., Nicklen, S., and Coulson, A.R. 1977. DNA sequencing with chain-terminating inhibitors. Proc. Natl. Acad. Sci. U.S.A. 74:5463-5467.
	Thomas, K.R. and Olivera, B.M. 1978. Processivity of DNA exonucleases. J. Biol. Chem. 253:424-429.
	Vales, L.D., Rabin, B.A., and Chase, J.W. 1982. Subunit structure of Escherichia coli exonuclease VII. J. Biol. Chem. 257:8799-8805.