On/off switch mediated by Exo+ polymerases: experimental analysis for its physiological and technological implications.
The potential physiological role and technological application of the premature termination of DNA polymerization through the off-switch of exo+ polymerases were studied using 3' phosphorothioate-modified or unmodified primers with single base mismatch distal to the 3' terminus. With exonuclease-digestible unmodified primers, a gradient premature termination of DNA polymerization was observed when amplified with exo+ polymerases. With 3' allele specific phosphorothioate-modified primers, an efficient off-switch effect occurred in the discrimination of a single nucleotide polymorphism when directly using genomic DNA. Clearly, the off-switch of exo+ polymerases is useful in biomedical research.
Zhang
Jia
J
Genomapping Inc., Tianjin, China.
Chen
Lin-Ling
LL
Guo
Zi-Fen
ZF
Peng
Cui-Ying
CY
Liao
Duan-Fang
DF
Li
Kai
K
D000483:Alleles; D001483:Base Sequence; D001704:Biopolymers; D004247:DNA / Q000737:chemistry / Q000378:metabolism; D017931:DNA Primers; D004259:DNA-Directed DNA Polymerase / Q000378:metabolism* / Q000502:physiology; D013873:Thionucleotides / Q000737:chemistry
D001704:Biopolymers; D017931:DNA Primers; D013873:Thionucleotides; D004247:DNA; D004259:DNA-Directed DNA Polymerase