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Carbohydrates and Nucleic Acids
Published in Michael B. Smith, A Q&A Approach to Organic Chemistry, 2020
A polynucleotide is a polymer of nucleosides linked together by phosphate linkages (usually monophosphate linkages). A polynucleotide of this type is called a nucleic acid. What is a deoxyribonucleotide?
Introduction to the Biological System
Published in Ashutosh Kumar Dubey, Amartya Mukhopadhyay, Bikramjit Basu, Interdisciplinary Engineering Sciences, 2020
Ashutosh Kumar Dubey, Amartya Mukhopadhyay, Bikramjit Basu
As shown in Figure 8.2, most of the DNA molecules are structurally characterized by two long polypeptide strands uniquely coiled around each other which form a double helix structure. Each of these strands, known as polynucleotides, are composed of four types of nucleotides, that is, nitrogen containing nucleobase, either cytosine (C), guanine (G), adenine (A), or thymine (T) as well as a sugar called deoxyribose and a phosphate group linked covalently into the polynucleotide chain with a sugar-phosphate backbone. This unique structural arrangement results in a characteristic sugar−phosphate backbone. The DNA structure is determined by the famous base pairing rules (A with T, and C with G). The complementary base pairing enables packing in energetically most favorable arrangement.
Biomolecular Processing and Molecular Electronics
Published in Sergey Edward Lyshevski, Molecular Electronics, Circuits, and Processing Platforms, 2018
Nucleic acids are polymers of monomers called nucleotides. Each nucleotide is itself composed of three parts, and a nitrogenous base is joined to a pentose that is bonded to a phosphate group. The DNA molecules consist of two polynucleotide chains (strands) that spiral around forming a double helix, which was discovered by Rosalind Franklin in 1952 through x-ray crystallography. These polynucleotide chains are held together by hydrogen bonds between the paired nitrogenous bases. DNA is a linear double-stranded polymer of the following four nucleotides (bases): Deoxyadenosine monophosphate or adenine (A)Deoxythymidine monophosphate or thymine (T) in DNA, and uracil (U) in RNADeoxyguanosine monophosphate or guanine (G)Deoxycytidine monophosphate or cytosine (C)
Cost-effective, high-yield production of Pyrobaculum calidifontis DNA polymerase for PCR application
Published in Preparative Biochemistry & Biotechnology, 2023
Kashif Maseh, Syed Farhat Ali, Shazeel Ahmad, Naeem Rashid
Assay for DNA polymerase activity was done as describe previously[14] by measuring the incorporation of TTP [methyl-3H] by using activated calf thymus DNA. The reaction mixture, in 20 µL, contained: 25 mM Tris–Cl pH 8.5, 4 mM MgCl2, 100 µM each of dATP, dGTP, dCTP, dTTP, 0.5 µCi TTP [methyl-3H] (78 Ci/mmol), 5 µg activated calf thymus DNA, 0.2 mg/ml BSA and 0.1% Tween 20. The reaction mixture was incubated at 75 °C for 5 min before enzyme addition. After adding the enzyme, aliquots were removed from the reaction mixture at various time intervals and spotted onto DE-81 filter paper disks (23 mm diameter, Whatman). The disks were air-dried, washed three times in sodium phosphate buffer pH 7.0 and finally washed with 70% ethanol. All washing steps were done for 2 min each. The filter paper disks were then dried and incorporated radioactivity on the filter disks was measured in counts per second (cps) by using Raytest Malisa scintillation counter (Berlin, Germany). One unit of DNA polymerase activity was defined as the amount of the enzyme required to incorporate 10 nmol [methyl 3H] TTP into a polynucleotide fraction (adsorbed on DE-81 filter disc) at 75 °C in 30 min.
Cloning, expression and characterization of a HER2-alpha luffin fusion protein in Escherichia coli
Published in Preparative Biochemistry and Biotechnology, 2019
Farzaneh Barkhordari, Nooshin Sohrabi, Fatemeh Davami, Fereidoun Mahboudi, Yeganeh Talebkhan Garoosi
Polynucleotide-adenosine glycosidase activity of the fusion protein was assessed on salmon sperm DNA (Sigma) according to the protocols.[29] In brief, different concentrations (3, 6, 9, and 12 µg) of the fusion and control proteins (unrelated scFv) were incubated in duplicate with 10 µg DNA at 30 °C for 40 min in a reaction buffer (50 mM magnesium acetate, 100 mM KCl, pH4.0) in a final volume of 300 µL. The reaction was stopped on ice. Polynucleotides were precipitated by ethanol and sodium acetate (salting out procedure) after keeping samples at –80 °C for 3 hr. The amount of the released bases into the supernatant was measured by Nanodrop (DeNovix) at 260 nm using the mixture of reaction buffer and alcohol/sodium acetate as the blank. The glycosidase activity of the fusion protein was assessed in comparison to the unrelated scFv (control) protein in triplicated tests.