Construction of chimera protein by using artificial restriction DNA cutter.
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.
Uehara
Akihiko
A
Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo, 153-8904, Japan.
Yamamoto
Yoji
Y
Watanabe
Akira
A
Aburatani
Hiroyuki
H
Komiyama
Makoto
M
D000818:Animals; D019556:COS Cells; D002563:Cerium; D002522:Chlorocebus aethiops; D004262:DNA Restriction Enzymes / Q000737:chemistry; D004274:DNA, Recombinant / Q000737:chemistry* / Q000378:metabolism; D003851:Deoxyribonucleases / Q000737:chemistry*; D004492:Edetic Acid / Q000737:chemistry; D049452:Green Fluorescent Proteins / Q000235:genetics; D006868:Hydrolysis; D010088:Oxidoreductases / Q000235:genetics; D020135:Peptide Nucleic Acids / Q000737:chemistry; D011993:Recombinant Fusion Proteins / Q000032:analysis / Q000235:genetics*
D004274:DNA, Recombinant; D020135:Peptide Nucleic Acids; D011993:Recombinant Fusion Proteins; C485184:enhanced green fluorescent protein; D049452:Green Fluorescent Proteins; D002563:Cerium; D004492:Edetic Acid; D010088:Oxidoreductases; D003851:Deoxyribonucleases; D004262:DNA Restriction Enzymes