scholarly journals Mononucleotide repeat expansions with non-natural polymerase substrates

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Alexander V. Chudinov ◽  
Vadim A. Vasiliskov ◽  
Viktoriya E. Kuznetsova ◽  
Sergey A. Lapa ◽  
Natalia A. Kolganova ◽  
...  
Keyword(s):  
Primer Extension ◽  
Current Paper ◽  
Mononucleotide Repeat ◽  
Biological Phenomenon ◽  
Primer Extension Reaction ◽  
Nucleotide Analogues ◽  
Full Size ◽  
Nucleoside Triphosphates ◽  
Repeat Expansions ◽  
Incorporation Efficiency

AbstractReplicative strand slippage is a biological phenomenon, ubiquitous among different organisms. However, slippage events are also relevant to non-natural replication models utilizing synthetic polymerase substrates. Strand slippage may notably affect the outcome of the primer extension reaction with repetitive templates in the presence of non-natural nucleoside triphosphates. In the current paper, we studied the ability of Taq, Vent (exo-), and Deep Vent (exo-) polymerases to produce truncated, full size, or expanded modified strands utilizing non-natural 2′-deoxyuridine nucleotide analogues and different variants of the homopolymer template. Our data suggest that the slippage of the primer strand is dependent on the duplex fluttering, incorporation efficiency for a particular polymerase-dNTP pair, rate of non-templated base addition, and presence of competing nucleotides.

scholarly journals Pyridone-based nucleotide analogues accepted for DNA biosynthesis

Biologija ◽  
2017 ◽  
Vol 63 (1) ◽  
Author(s):  
Algirdas Mikalkėnas ◽  
Bazilė Ravoitytė ◽  
Daiva Tauraitė ◽  
Saulius Serva
Keyword(s):  
Dna Synthesis ◽  
Dna Polymerases ◽  
Primer Extension ◽  
Nucleotide Analogues ◽  
Dna Biosynthesis ◽  
Nucleoside Triphosphates

Novel pyridone-based nucleotides were investigated for maintenance of DNA synthesis, carried on by representatives of different classes of polymerases. Five different nucleoside triphosphates of similar structure were addressed, demonstrating involvement in primer extension. The  nucleotides containing 4-chloro- and 4-bromo-2-pyridone as a nucleobase were preferred by all DNA polymerases tested. These nucleotides were incorporated up to several instances in a  row and unexpectedly failed to ensure DNA primer termination. Temporary polymerase stalling obeys kinetic constrain and is alleviated by availability of nucleotides readily acquired by the polymerase. Lack of inhibitory properties of 2-pyridone nucleobase confirms the processivity of DNA polymerases challenged by incorporated nucleotides.

Multi-allele DNA biosensor for the rapid genotyping of ‘nondeletion’ alpha thalassaemia mutations in HBA1 and HBA2 genes by means of multiplex primer extension reaction

2015 ◽  
Vol 446 ◽  
pp. 241-247 ◽  
Author(s):  
Margarita Petropoulou ◽  
Amalia Poula ◽  
Jan Traeger-Synodinos ◽  
Emmanuel Kanavakis ◽  
Theodore K. Christopoulos ◽  
...  
Keyword(s):  
Dna Biosensor ◽  
Primer Extension ◽  
Primer Extension Reaction ◽  
Alpha Thalassaemia

scholarly journals Mutant T7 RNA polymerase is capable of catalyzing DNA primer extension reaction

FEBS Letters ◽  
1998 ◽  
Vol 423 (2) ◽  
pp. 189-192 ◽  
Author(s):  
E.E Rusakova ◽  
V.L Tunitskaya ◽  
L.V Memelova ◽  
S.V Kochetkova ◽  
D.A Kostyuk ◽  
...  
Keyword(s):  
Rna Polymerase ◽  
Primer Extension ◽  
T7 Rna Polymerase ◽  
Primer Extension Reaction

Primer Extension Reaction Assays for Incorporation of Deoxynucleotide Analogue into DNA

2015 ◽  
Vol 33 (2) ◽  
pp. 192-198
Author(s):  
Xilong Wang ◽  
Xun Chen ◽  
Yonghui Liu ◽  
Jin Zhu
Keyword(s):  
Primer Extension ◽  
Primer Extension Reaction
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