Skip Navigation


Journal of Biochemistry Advance Access originally published online on August 24, 2007
Journal of Biochemistry 2007 142(2):283-292; doi:10.1093/jb/mvm137
This Article
Right arrow Full Text
Right arrow Full Text (PDF)
Right arrow All Versions of this Article:
142/2/283    most recent
mvm137v1
Right arrow Alert me when this article is cited
Right arrow Alert me if a correction is posted
Services
Right arrow Email this article to a friend
Right arrow Similar articles in this journal
Right arrow Similar articles in PubMed
Right arrow Alert me to new issues of the journal
Right arrow Add to My Personal Archive
Right arrow Download to citation manager
Right arrow Request Permissions
Google Scholar
Right arrow Articles by Uemori, T.
Right arrow Articles by Kato, I.
Right arrow Search for Related Content
PubMed
Right arrow PubMed Citation
Right arrow Articles by Uemori, T.
Right arrow Articles by Kato, I.
Social Bookmarking
Add to CiteULike   Add to Connotea   Add to Del.icio.us  
What's this?

© 2007 The Japanese Biochemical Society.

Investigation of the Molecular Mechanism of ICAN, a Novel Gene Amplification Method

Takashi Uemori1, Hiroyuki Mukai2, Osamu Takeda2, Mariko Moriyama1,{dagger}, Yoshimi Sato1, Shigekazu Hokazono1, Nariaki Takatsu2, Kiyozo Asada1,* and Ikunoshin Kato1

1Biotechnology Research Laboratories, Takara Bio Inc., Seta 3-4-1 Otsu, Shiga 520-2193, Japan; and 2Products Development Center, Takara Bio Inc., 2257, Noji, Kusatsu, Shiga 525-0055, Japan

*To whom correspondence should be addressed. Tel: +81 77-543-7234, Fax: +81 77-543-7295, E-mail: asadak{at}takara-bio.co.jp

Received April 3, 2007; Accepted June 12, 2007


  Abstract

Isothermal and Chimeric primer-initiated Amplification of Nucleic acids (ICAN) allows the amplification of target DNA under isothermal conditions at around 55°C using only a pair of 5'-DNA-RNA-3' chimeric primers, thermostable RNaseH and a DNA polymerase with strand-displacing activity (H. Mukai et al. J. Biochemistry, in the preceding paper in this issue). Here we elucidated the mechanism of ICAN by analysing the nicking site of RNaseH, behaviour of chimeric primers and extension products. We found that the ICAN reaction was composed of two unique mechanisms, multi-priming and template-switching, that were responsible for the highly efficient amplifying capability of ICAN. The simultaneous occurrence of two types of reactions, one based on multi-priming and the other based on template-switching, is likely to drive the DNA amplification in ICAN.

Key Words: ICAN, multi-priming, RNaseH, strand displacement, template-switching

Abbreviations: FITC, fluorescein isothiocianate; ICAN, isothermal and chimeric primer-initiated amplification of nucleic acids; ROX, 6-carboxy-X-rhodamine; Tli RNaseH, Thermococcus litoralis RNaseH; XRITC, X-rhodamine isothiocyanate


Add to CiteULike CiteULike   Add to Connotea Connotea   Add to Del.icio.us Del.icio.us    What's this?




Disclaimer:
Please note that abstracts for content published before 1996 were created through digital scanning and may therefore not exactly replicate the text of the original print issues. All efforts have been made to ensure accuracy, but the Publisher will not be held responsible for any remaining inaccuracies. If you require any further clarification, please contact our Customer Services Department.