ABB  Vol.4 No.4 , April 2013
Nickase-dependent isothermal DNA amplification
Author(s) Yan He, Tao Jiang*
ABSTRACT

We developed a nicking endonuclease dependent DNA amplification (NDA), using Nt.BstNBI to catalyze single-stranded nick on double-stranded DNA, and Bst DNA polymerase to make extension while sealing the nick and displacing the downstream strand. The displaced single-stranded DNA thereby serves as template for primers hybridization and extension, resulting in exponential synthesis of target DNA under isothermal condition. Over 105 folds target DNA amplification can be achieved in 30 minutes, generating DNA product suitable for both diagnosis and DNA cloning. This NDA strategy does not require thermal cycling or prerequisite nucleotides modification, making it suitable for application in the field and at the point-of-care.


Cite this paper
He, Y. and Jiang, T. (2013) Nickase-dependent isothermal DNA amplification. Advances in Bioscience and Biotechnology, 4, 539-542. doi: 10.4236/abb.2013.44070.
References
[1]   Saiki, R.K., Gelfand, D.H., Stoffel, S., Scharf, S.J., Higuchi, R., Horn, G.T., Mullis, K.B. and Erlich, H.A. (1988) Primer-directed enzymatic amplification of DNA with a thermostable DNA polymerase. Science, 239, 487491. doi:10.1126/science.2448875

[2]   Taylor, G.R. and Robinson, P. (1998) The polymerase chain reaction, from functional genomics to high-school practical classes. Current Opinion Biotechnology, 9, 3542. doi:10.1016/S0958-1669(98)80081-5

[3]   Gill, P. and Ghaemi, A. (2008) Nucleic acid isothermal amplification technologies—A review. Nucleosides Nucleotides Nucleic Acids, 27, 224-243. doi:10.1080/15257770701845204

[4]   Walker, G.T., Little, M.C., Nadeau, J.G. and Shank, D.D. (1992) Isothermal in vitro amplification of DNA by a restriction enzyme/DNA polymerase system. Proceedings of National Academic Science of the USA, 89, 392-396. doi:10.1073/pnas.89.1.392

[5]   Walker, G.T., Fraiser, M.S., Schram, J.L., Little, M.C., Nadeau, J.G. and Malinowski, D.P. (1992) Strand displacement amplification—An isothermal, in vitro DNA amplification technique. Nucleic Acids Research, 20, 16911696. doi:10.1093/nar/20.7.1691

[6]   Walker, G.T. (1993) Empirical aspects of strand displacement amplification. PCR Methods Application, 3, 1-6. doi:10.1101/gr.3.1.1

[7]   Notomi, T., Okayama, H., Masubuchi, H., Yonekawa, T., Watanabe, K., Amino, N. and Hase, T. (2000) Loop-mediated isothermal amplification of DNA. Nucleic Acids Research, 28, e63. doi:10.1093/nar/28.12.e63

[8]   Vincent, M., Xu, Y. and Kong, H. (2004) Helicase-dependent isothermal DNA amplification. EMBO Rep, 5, 795800. doi:10.1038/sj.embor.7400200

[9]   An, L., Tang, W., Ranalli, T.A., Kim, H.J., Wytiaz, J. and Kong, H. (2005) Characterization of a thermostable UvrD helicase and its participation in helicase-dependent amplification. Journal of Biological Chemistry, 280, 2895228958. doi:10.1074/jbc.M503096200

[10]   Kong, H., Higgins, L., Dalton, M., Kucera, R. and Schildkraut, I. (2001) Cloning and producing the N.BstNBI nicking endonuclease. US Patent No. 6191267.

[11]   Chan, S.H., Zhu, Z., Van Etten, J.L. and Xu, S.Y. (2004) Cloning of CviPII nicking and modification system from chlorella virus NYs-1 and application of Nt.CviPII in random DNA amplification. Nucleic Acids Research, 32, 6187-6199. doi:10.1093/nar/gkh958

[12]   Van Ness, J., Galas, D. and Van Ness, L. (2006) Nucleic acid amplification using nicking agents. US Patent No. 7112423.

[13]   Kucera, R. (2007) Genome amplification. International Patent No. WO/2007/056173.

[14]   Yao, Z. and Lidgard, G. (2008) Method for rapid, singlestep strand displacement amplification of nucleic acids. International Patent No. WO/2008/066979.

[15]   Zyrina, N.V., Zheleznaya, L.A., Dvoretsky, E.V., Vasiliev, V.D., Chernov, A. and Matvienko, N.I. (2007) N.BspD6I DNA nickase strongly stimulates template-independent synthesis of non-palindromic repetitive DNA by Bst DNA polymerase. Biological Chemistry, 388, 367-372. doi:10.1515/BC.2007.043

 
 
Top