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Nucleic Acids Symposium Series 2005 49(1):279-280; doi:10.1093/nass/49.1.279
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© 2005 Oxford University Press

Construction of chimera protein by using artificial restriction DNA cutter

Akihiko Uehara, Yoji Yamamoto, Akira Watanabe, Hiroyuki Aburatani and Makoto Komiyama

Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo, 153-8904, Japan

We have already developed artificial restriction DNA cutter (ARCUT), which can hydrolyze double-stranded DNA site-selectively, by using Ce(IV)/EDTA in combination with two pseudo-complementary peptide nucleic acids (pcPNAs). Here, ARCUT was used to prepare a chimera protein. The gene for WW-domain containing oxidoreductase (WWOX) was clipped off by ARCUT just before its stop codon, and ligated with the gene for enhanced green fluorescent protein (EGFP). Conventional PCR for insertion of restriction enzyme site is never required.


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