Gene manipulation using artificial restriction DNA cutter.
Artificial restriction DNA cutter (ARCUT), which we developed recently, was used for manipulation of plasmid DNA. PCR product was inserted into pBR322 plasmid vector using ARCUT, and E. coli cells were transfected with this recombinant plasmid DNA. Successful growth of cells shows that the recombination proceeds without any unexpected mutations. Furthermore, this artificial system was applied to PCR-free construction of chimera protein.
Yamamoto
Yoji
Y
Research Center for Advanced Science and Technology, The University of Tokyo, 4-6-1 Komaba, Meguro-ku, Tokyo 153-8904, Japan.
Zhou
Jing-Min
JM
Tomita
Takafumi
T
Uehara
Akihiko
A
Aiba
Yuichiro
Y
Watanabe
Akira
A
Aburatani
Hiroyuki
H
Komiyama
Makoto
M
D002563:Cerium / Q000737:chemistry; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics; D005822:Genetic Vectors; D006868:Hydrolysis; D020135:Peptide Nucleic Acids / Q000737:chemistry; D010957:Plasmids / Q000235:genetics*; D016133:Polymerase Chain Reaction; D015202:Protein Engineering / Q000379:methods*; D011993:Recombinant Fusion Proteins / Q000235:genetics*; D011995:Recombination, Genetic; D014162:Transfection
D020135:Peptide Nucleic Acids; D011993:Recombinant Fusion Proteins; D002563:Cerium; D004247:DNA