The structural coordinates for human DNA polymerase
[alpha] in ternary complex with RNA-primed DNA template and aphidicolin were obtained from the PDB-ID: 4Q5V with 2.50 A resolutions.
Reactions that reached exhaustion within 30 min were calibrated by normalizing the minimum fluorescent value of each progress-curve to zero, and the maximum fluorescent value to the average number of extendable bases per polymerase
. Then, the initial slope of the curve corresponds to the average initial rate in nucleotides per second polymerase
Based on the 2.5 kb gene sequence information of Taq DNA polymerase
gene (Di et al., 2012), a pair of primers (Taq1: 5'-GGCCGAATTCATGCTGCCCCTCTTTGAGCCC-3') and Taq2: 5'-GGCCGAATTCTCACTCCTTGGCGGAGAGCC-3') were designed, with EcoRI located at each end of the gene.
chain reaction (PCR)-based techniques have been developed for routine clinical malaria diagnosis, e.g.
This study has confirmed the existence of HBV DNA polymerase
resistance mutations for NAs in 24 (8.9%) of 269 treatment-naive patients, composed of five primary drug resistance mutations (rtT184G, rtS202I, rtM204I/V, rtN236T, and rtM250V), one secondary resistance mutation (rtL180M), and one codon mutation that might be baseline polymorphisms without any clinical significance (rtV207I).
As it can be seen from the results of electrophoresis, Taq polymerase
is capable to effectively incorporate in the sequence of a strand only modified nucleotide 5-(C8 Alkyne)-2'-Deoxyuridine-5'-Triphosphate.
Previous studies have shown that PB1-F2 of the A/WSN/33 strain (WSN;H1N1) upregulates viral polymerase
activity in vitro .
Therefore, the methods for rescuing virus in vivo with high efficiency and no need for RNA preparation in vitro are urgently established.T7 RNAP, one of single-subunit RNA polymerases
, is widely employed to generate high specific activity labeled RNA probes, RNA for in vitro translation, biologically active mRNA, and preparative quantities of defined length RNA by the run off transcription method (Schenborn and Mierendorf, 1985).
In vivo DNA replication methods for most organisms rely on a class of enzymes known as DNA helicases, which act to locally unwind dsDNA to create small ssDNA regions suitable for polymerase
One could suppose that stabilization of the beta-structured state of PB1 N-terminus would result in inhibition of the interaction of this protein with PA, polymerase
complex assembly, and reproduction of viral RNA.
By direct sequencing of clones from two independent PCR experiments with Taq polymerase
, we observed 99 unique mutations out of >100 kbp of target DNA sequence.
Assay for Incorporation opposite 5' -Neighboring Bases of G or Oz.