Download or read book Polymerase Activity of Chimeric Polymerase written by Wing-hong Chin and published by . This book was released on 2012 with total page 194 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Polymerase Activity of Chimeric Polymerase written by Wing-Hong Chin and published by Open Dissertation Press. This book was released on 2017-01-26 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt: This dissertation, "Polymerase Activity of Chimeric Polymerase: a Determining Factor for an Influenza Virus to Be a Pandemic Strain" by Wing-hong, Chin, 錢永康, was obtained from The University of Hong Kong (Pokfulam, Hong Kong) and is being sold pursuant to Creative Commons: Attribution 3.0 Hong Kong License. The content of this dissertation has not been altered in any way. We have altered the formatting in order to facilitate the ease of printing and reading of the dissertation. All rights not granted by the above license are retained by the author. Abstract: The influenza polymerase is a complex of three subunits, polymerase basic protein 2 (PB2), polymerase basic protein 1 (PB1) and polymerase acidic protein (PA). It associates with the viral RNA segment and nucleoprotein (NP) to form a viral ribonucleoprotein (vRNP) complex which is important for transcription and replication of the viral genome. Concurrently, the previous three influenza pandemics viruses contain reassorted vRNP of different origins. This leads to the aim of study to investigate the role of polymerase in the pandemic viruses. By reconstitution of vRNPs in human cells, it was demonstrated that vRNPs of H2N2 and H3N2 pandemic viruses had higher polymerase activity than the H2N2 seasonal viruses in-between them. The recombinant virus with H2N2 pandemic vRNP also showed faster growth kinetics in the early stage of viral replication and better adaptability to the selective environment with neuraminidase inhibitor than the recombinant virus with H2N2 seasonal vRNP, which had a lower polymerase activity. Reconstitution of chimeric vRNPs of H2N2 pandemic and seasonal viruses revealed that PB2, PB1 and PA were responsible for the difference in polymerase activity between them. Five residues, one in PB2, three in PB1 and one in PA were identified to be significant for the polymerase activity change. These polymerase subunits and residues may act as part of the determining factors for the H2N2 pandemic virus. Furthermore, PB2-627 has been shown to have stringent host specificity and affect polymerase activity and viral replication. Recombinant viruses in mammalian and avian cells with random mutation were generated at this position. It showed that the amino acids at this position are not restricted to those appear in the nature for generating viable viruses. It was also observed that the avian-derived viruses generally had lower polymerase activity and reduced growth kinetics in mammalian cells, while part of the mammalian-derived viruses had lower polymerase activity and reduced growth kinetics in avian cells. This consolidated the role of PB2-627 on host specificity and demonstrated the possibility of some novel amino acids for this position, which may play a role in the future influenza pandemic. The 2009 H1N1 pandemic virus contains a reassorted vRNP with subunits of avian, human and swine origins. This prompts me to compare the polymerase activity of all the 81 possible combinations of chimeric vRNPs of three different origins. The results were statistically analyzed and several single subunit factors and interactions between vRNP subunits were identified to significantly affect the polymerase activity. In order to reduce the effort and resources required, a fractional factorial design of 27 experimental runs was developed to substitute the 81-combination full factorial design for identifying the significant single subunit factors that affect the polymerase activity. Overall, this study identified some factors that may contribute to a pandemic virus and allows us to have better understanding of the role of polymerase in a pandemic virus. These findings may contribute to evaluating the pandemic potential of the novel virus that emerges or may emerge in the nature and enhances the preparedness towards the next pandemic influenza. DOI: 10.5353/th_b4979918 Subjects: RNA polymerases Influenza viruses Nucleoproteins
Download or read book Polymerase Activity of Chimeric Polymerase written by Wing-hong Chin and published by . This book was released on 2012 with total page 0 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book DNA polymerases in Biotechnology written by Zvi Kelman and published by Frontiers Media SA. This book was released on 2015-03-18 with total page 147 pages. Available in PDF, EPUB and Kindle. Book excerpt: DNA polymerases are core tools for molecular biology including PCR, whole genome amplification, DNA sequencing and genotyping. Research has focused on discovery of novel DNA polymerases, characterization of DNA polymerase biochemistry and development of new replication assays. These studies have accelerated DNA polymerase engineering for biotechnology. For example, DNA polymerases have been engineered for increased speed and fidelity in PCR while lowering amplification sequence bias. Inhibitor resistant DNA polymerase variants enable PCR directly from tissue (i.e. blood). Design of DNA polymerases that efficiently incorporate modified nucleotide have been critical for development of next generation DNA sequencing, synthetic biology and other labeling and detection technologies. The Frontiers in Microbiology Research Topic on DNA polymerases in Biotechnology aims to capture current research on DNA polymerases and their use in emerging technologies.
Download or read book DNA Directed DNA Polymerases Evolve From Reverse Transcriptase written by Lei Zhang and published by . This book was released on 2013 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Differential Effect of N Carboxymethylisatoylation on the DNA Polymerase Activity the 5 3 exonuclease Activity and the 3 5 exonuclease Activity of DNA Polymerase I of Escherichia Coli written by Hans Klenow and published by . This book was released on 1981 with total page 7 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Chimeric Dinucleotides written by Michael G Mohsen and published by . This book was released on 2020 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Industrial Enzymes written by Julio Polaina and published by Springer Science & Business Media. This book was released on 2007-05-16 with total page 629 pages. Available in PDF, EPUB and Kindle. Book excerpt: Recent developments in genetic engineering and protein chemistry are bringing ever more powerful means of analysis to bear on the study of enzyme structure. This volume reviews the most important types of industrial enzymes. In a balanced manner it covers three interrelated aspects of paramount importance for enzyme performance: three-dimensional protein structure, physicochemical and catalytic properties, and the range of both classical and novel applications.
Download or read book The Polymerase Chain Reaction written by Kary B. Mullis and published by Birkhauser. This book was released on 1994 with total page 490 pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book DNA Polymerase Activity in Proliferating and Antigen stimulated Tissues written by Maria Giulia Battelli and published by . This book was released on 1979 with total page pages. Available in PDF, EPUB and Kindle. Book excerpt:
Download or read book Functions of the DNA Polymerase Delta Replicase in Lagging Strand Replication written by Joseph L. Stodola and published by . This book was released on 2016 with total page 178 pages. Available in PDF, EPUB and Kindle. Book excerpt: The work described in this dissertation focuses on several aspects of DNA replication in the model organism Saccharomyces cerevisiae, with particular attention paid to the function of the replicative DNA polymerase [delta] (Pol [delta]), and its functions in Okazaki fragment synthesis and maturation. The first major theme of this dissertation is investigating the role that metal binding motifs play in the structure and function of Pol [delta] and other budding yeast polymerases. First, I discuss the role that two metal binding motifs within the catalytic subunit of Pol [delta] play in creating the multi-subunit polymerase complex and in promoting crucial interactions with the replication sliding clamp, proliferating cell nuclear antigen (PCNA). Next, I describe work defining the importance of similar metal binding motifs in the translesion DNA polymerase [zeta] (Pol [zeta]). This yielded the observation that the two accessory subunits of Pol [delta], Pol31 and Pol32, are also constitutive members of a four-subunit Pol [zeta] complex. Finally, I describe the creation of a chimeric DNA polymerase comprising the bacteriophage RB69 DNA polymerase fused to the metal binding domain of the Pol [delta] catalytic subunit. We show that this chimeric polymerase can form a multimeric complex containing the Pol [delta] accessory subunits, interact with PCNA, and support DNA replication in vivo. This data provided insight into the structural requirements of the lagging strand replication machinery. The second major theme is Pol [delta]'s crucial role in synthesizing Okazaki fragments and participating in the removal of initiator RNA, called Okazaki fragment maturation. I first describe my work developing a system to study the activity of Pol [delta] in higher kinetic detail than previous studies, using rapid-quench techniques. This work yielded insights into how Pol [delta] performs DNA synthesis and strand displacement synthesis, as well as accomplishes nick translation, requiring collaboration between Pol [delta] and the flap-endonuclease FEN1. The next chapter describes the production and characterization of engineered PCNA heterotrimers. These proteins were produced to test the 'toolbelt' model, which is the hypothesis that PCNA binds multiple enzymes simultaneously to increase the efficiency of DNA metabolism processes involving multiple enzymes. Finally, there has been a growing interest among those studying lagging strand synthesis into how potential impediments to the Okazaki fragment maturation machinery are resolved; we show that although the transcription factor Rap1 can block strand displacement synthesis by Pol [delta] when it is bound to DNA, this block can be resolved through the action of the helicase Pif1. In sum, these studies provide insight into how Pol [delta]'s structure dictates its function, as well as addresses larger mechanistic questions concerning how lagging strand DNA replication is accomplished.
Download or read book Protein Engineering Handbook Volume 3 written by Stefan Lutz and published by John Wiley & Sons. This book was released on 2012-09-14 with total page 723 pages. Available in PDF, EPUB and Kindle. Book excerpt: This introduction collects 17 innovative approaches to engineer novel and improved proteins for diverse applications in biotechnology, chemistry, bioanalytics and medicine. As such, key developments covered in this reference and handbook include de novo enzyme design, cofactor design and metalloenzymes, extremophile proteins, and chemically resistant proteins for industrial processes. The editors integrate academic innovations and industrial applications so as to arrive at a balanced view of this multi-faceted topic. Throughout, the content is chosen to complement and extend the previously published two-volume handbook by the same editors, resulting in a superb overview of this burgeoning field.
Download or read book Virus Protein and Nucleoprotein Complexes written by J. Robin Harris and published by Springer. This book was released on 2018-06-13 with total page 443 pages. Available in PDF, EPUB and Kindle. Book excerpt: The Subcellular Biochemistry series has recently embarked upon an almost encyclopaedic coverage of topics relating to the structure and function of macromolecular complexes (Volumes 82, 83 and 87). The present multi-author text covers numerous aspects of current research into molecular virology, with emphasis upon viral protein and nucleoprotein structure and function. Structural data from cryo-electron microscopy and X-ray crystallography is displayed throughout the book. The 17 chapters in the book cover diverse interesting topics, all currently under investigation, contributed by authors who are active actively involved in present-day research. Whilst structural aspects predominate, there is much consideration of the structure-function relationship. In addition, the book correlates with and extends from Volume 68 of the series “Structure and Physics of Viruses: An Integrated Textbook”. This book is directed primarily at professionals that work in the broad field of Structural Biology and will be of particular interest to Structural Virologists. The editors, David Bhella and Robin Harris, have much experience in virology and protein structure, respectively. Dr Bhella is Director of the Scottish Macromolecular Imaging Centre. Professor Robin Harris is the long-standing Series Editor of the Subcellular Biochemistry series. He has edited and contributed to several books in the series.
Download or read book Protein nucleic Acid Interaction written by Wolfram Saenger and published by . This book was released on 1989 with total page 230 pages. Available in PDF, EPUB and Kindle. Book excerpt: This volume contains a series of essays which describe a range of problems in the field of nucleic-acid interactions, investigated by a variety of techniques. An introductory chapter on DNA-protein interactions in the regulation of gene expression is followed by papers on selected model systems.
Download or read book Modified Nucleic Acids written by Kazuhiko Nakatani and published by Springer. This book was released on 2016-04-04 with total page 280 pages. Available in PDF, EPUB and Kindle. Book excerpt: This book spans diverse aspects of modified nucleic acids, from chemical synthesis and spectroscopy to in vivo applications, and highlights studies on chemical modifications of the backbone and nucleobases. Topics discussed include fluorescent pyrimidine and purine analogs, enzymatic approaches to the preparation of modified nucleic acids, emission and electron paramagnetic resonance (EPR) spectroscopy for studying nucleic acid structure and dynamics, non-covalent binding of low- and high-MW ligands to nucleic acids and the design of unnatural base pairs. This unique book addresses new developments and is designed for graduate level and professional research purposes.
Download or read book Dna Polymerases Discovery Characterization And Functions In Cellular Dna Transactions written by Ulrich Hubscher and published by World Scientific. This book was released on 2010-04-14 with total page 338 pages. Available in PDF, EPUB and Kindle. Book excerpt: Maintenance of the information embedded in the genomic DNA sequence is essential for life. DNA polymerases play pivotal roles in the complex processes that maintain genetic integrity. Besides their tasks in vivo, DNA polymerases are the workhorses in numerous biotechnology applications such as the polymerase chain reaction (PCR), cDNA cloning, genome sequencing, nucleic acids-based diagnostics and in techniques to analyze ancient and otherwise damaged DNA. Moreover, some diseases are related to DNA polymerase defects, and chemotherapy through inhibition of DNA polymerases is used to fight HIV, Herpes and Hepatitis B and C infections. We have recently witnessed the discovery of an abundance of novel DNA polymerases in viruses, bacteria, archaea and eukaryotes with specialized properties whose physiological functions are only beginning to be understood. This book summarizes the current knowledge of these fascinating enzymes. It is intended for a wide audience from basic scientists, to diagnostic laboratories and to clinicians who seek a better understanding of these fascinating enzymes.
Download or read book Applications of Chimeric Genes and Hybrid Proteins Part C Protein Protein Interactions and Genomics written by and published by Elsevier. This book was released on 2000-10-28 with total page 701 pages. Available in PDF, EPUB and Kindle. Book excerpt: The critically acclaimed laboratory standard for more than forty years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Since 1955, each volume has been eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with more than 300 volumes (all of them still in print), the series contains much material still relevant today--truly an essential publication for researchers in all fields of life sciences.
