Development of an unnatural base pair for efficient PCR amplification.
An unnatural base pair system could expand the genetic alphabet, enabling the site-specific incorporation of extra, functional components into nucleic acids and proteins. We developed an unnatural base pair between 7-(2-thienyl)imidazo[4,5-b]pyridine (denoted by Ds) and 2-nitropyrrole (denoted by Pn), which specifically and efficiently functions in DNA amplification by PCR. After 20 cycles of PCR, the mutation rate of the Ds-Pn pair in an amplified DNA fragment was approximately 1%. The Ds-Pn pair system in combination with other unnatural base pairs could be useful for DNA/RNA-based biotechnology.
Hirao
Ichiro
I
Protein Research Group, RIKEN Genomic Sciences Center, 1-7-22 Suehiro-cho, Tsurumi-ku, Yokohama, Kanagawa 230-0045, Japan. ihirao@riken.jp
Mitsui
Tsuneo
T
Kimoto
Michiko
M
Yokoyama
Shigeyuki
S
D020029:Base Pairing; D005815:Genetic Code; D005818:Genetic Engineering; D007093:Imidazoles / Q000737:chemistry*; D016133:Polymerase Chain Reaction; D011725:Pyridines / Q000737:chemistry*; D011758:Pyrroles / Q000737:chemistry*
C528342:2-nitropyrrole; C528341:7-(2-thienyl)imidazo(4,5-b)pyridine; D007093:Imidazoles; D011725:Pyridines; D011758:Pyrroles