Abstract
Bacterial NAD-dependent Taq and Tth DNA ligases are capable of significantly increasing the yield of long PCR products when the amplification is carried out using bacterial family A DNA polymerases, e.g. Taq or Tth DNA polymerases, or with enzymatic blends containing these polymerases. We also show that Taq and Tth DNA ligases improve the results of PCR in the absence of NAD and therefore in the absence of DNA ligase activity. These observations suggest that bacterial DNA ligases can interact with these DNA polymerases, presumably as accessory proteins, thereby enhancing the efficiency of DNA polymerization.
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References
Barnes, W. M. (1994) Proc. Natl. Acad. Sci. USA, 91, 2216–2220.
Cheng, S., Fockler, C., Barnes, W. M., and Higuchi, R. (1994) Proc. Natl. Acad. Sci. USA, 91, 5695–5699.
Barnes, W. M. (1992) Gene, 112, 29–35.
Fromenty, B., Demeilliers, C., Mansouri, A., and Pessayre, D. (2000) Nucleic Acids Res., 28, e50.
Hogrefe, H. H., Hansen, C. J., Scott, B. R., and Nielson, K. B. (2002) Proc. Natl. Acad. Sci. USA, 99, 596–601.
Fogg, M. J., Pearl, L. H., and Connolly, B. A. (2002) Nature Struct. Biol., 9, 922–927.
Motz, M., Kober, I., Girardot, C., Loeser, E., Bauer, U., Albers, M., Moeckel, G., Minch, E., Voss, H., Kilger, C., and Koegl, M. (2002) J. Biol. Chem., 277, 16179–16188.
Prelich, G., and Sillman, B. (1988) Cell, 53, 117–126.
Cann, I. K. O., Ishino, S., Hayashi, I., Komori, K., Toh, H., Morikawa, K., and Ishino, Y. (1999) J. Bacteriol., 181, 6591–6599.
Bullard, J. M., Williams, J. C., Acker, W. K., Jacobi, C., Janjc, N., and McHenry, C. S. (2002) J. Biol. Chem., 277, 13401–13408.
Modrich, P., and Richardson, C. C. (1975) J. Biol. Chem., 250, 5508–5514.
Takahashi, M., Yamaguchi, E., and Uchida, T. (1984) J. Biol. Chem., 259, 10041–10047.
Barany, F., and Gelfand, D. H. (1991) Gene, 109, 1–11.
Kornberg, A., and Baker, T. A. (1992) DNA Replication, 2nd Edn., W. H. Freeman and Company, New York.
Lauer, G., Rudd, E. A., McKay, D. L., Ally, A., Ally, D., and Backman, K. C. (1991) J. Bacteriol., 173, 5047–5053.
Billen, D., and Hellermann, G. R. (1976) J. Bacteriol., 126, 785–793.
Sakakibara, Y. (1978) J. Mol. Biol., 124, 373–389.
Kornberg, A. (1974) DNA Synthesis, W. H. Freeman and Company, San Francisco.
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Published in Russian in Biokhimiya, 2009, Vol. 74, No. 5, pp. 685–690.
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Ignatov, K.B., Kramarov, V.M. DNA ligases from thermophilic bacteria enhance PCR amplification of long DNA sequences. Biochemistry Moscow 74, 557–561 (2009). https://doi.org/10.1134/S0006297909050113
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DOI: https://doi.org/10.1134/S0006297909050113