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Systematic Analysis of Enzymatic DNA Polymerization Using Oligo DNA Templates and Triphosphate Analogs Involving 2’, 4’-Bridged Nucleosides


Affiliations
1 Department of Pharmacy, Nizam Institute of Pharmacy, Deshmukhi (V), Pochampally (M), Behind Mount Opera, Yadadri (Dist)-508284, Telangana, India
2 Department of Pharmaceutics, Nizam Institute of Pharmacy, Deshmukhi (V), Pochampally (M), Behind Mount Opera, Yadadri (Dist)-508284, Telangana, India
3 Department of Pharmaceutical Analysis and Quality Assurance, Nizam Institute of Pharmacy, Deshmukhi (V), Pochampally (M), Behind Mount Opera, Yadadri (Dist)-508284, Telangana, India
     

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In order to systematically analyze the effects of nucleosides modification of sugar in DNA polymerase reaction, we synthesized 16 modified templates containing 2', 4'-Bridged nucleoside-5'-triphosphates with different bridged structures. Among five types of thermos table DNA polymerases used, Taq, Phusion HF, vent (exo-), KOD Dash and KOD (exo-). The KOD Dash and KOD (exo-) DNA with polymerizes could smoothly read through modified templates containing 2'-0,4'-C-methylene linked nucleotides at standard enzyme concentration for five minutes. The modified templates containing sequence of seven successive 2',4'-bridged nucleotides could not be completely transcribed by any of the DNA polymerase used, yields of longer elongated products decreased in order to steric bulkiness of the modified sugar. Successive incorporation of 2',4'-bridged nucleotides into extending strands using 2',4'-bridged nucleotide -5'-triphosphate was much more difficult.

Keywords

Nucleosides, Coding, Polymerization, Systematic Analysis.
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  • Systematic Analysis of Enzymatic DNA Polymerization Using Oligo DNA Templates and Triphosphate Analogs Involving 2’, 4’-Bridged Nucleosides

Abstract Views: 41  |  PDF Views: 1

Authors

Ayesha Begum
Department of Pharmacy, Nizam Institute of Pharmacy, Deshmukhi (V), Pochampally (M), Behind Mount Opera, Yadadri (Dist)-508284, Telangana, India
Syeda Sharifa Nida
Department of Pharmacy, Nizam Institute of Pharmacy, Deshmukhi (V), Pochampally (M), Behind Mount Opera, Yadadri (Dist)-508284, Telangana, India
Nuha Rasheed
Department of Pharmaceutics, Nizam Institute of Pharmacy, Deshmukhi (V), Pochampally (M), Behind Mount Opera, Yadadri (Dist)-508284, Telangana, India
Abdul Saleem Mohammad
Department of Pharmaceutical Analysis and Quality Assurance, Nizam Institute of Pharmacy, Deshmukhi (V), Pochampally (M), Behind Mount Opera, Yadadri (Dist)-508284, Telangana, India

Abstract


In order to systematically analyze the effects of nucleosides modification of sugar in DNA polymerase reaction, we synthesized 16 modified templates containing 2', 4'-Bridged nucleoside-5'-triphosphates with different bridged structures. Among five types of thermos table DNA polymerases used, Taq, Phusion HF, vent (exo-), KOD Dash and KOD (exo-). The KOD Dash and KOD (exo-) DNA with polymerizes could smoothly read through modified templates containing 2'-0,4'-C-methylene linked nucleotides at standard enzyme concentration for five minutes. The modified templates containing sequence of seven successive 2',4'-bridged nucleotides could not be completely transcribed by any of the DNA polymerase used, yields of longer elongated products decreased in order to steric bulkiness of the modified sugar. Successive incorporation of 2',4'-bridged nucleotides into extending strands using 2',4'-bridged nucleotide -5'-triphosphate was much more difficult.

Keywords


Nucleosides, Coding, Polymerization, Systematic Analysis.