Chan, Wai Hon. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2010. / Includes bibliographical references (leaves 134-141). / Abstracts in English and Chinese. / Acknowledgements --- p.2 / Abstract --- p.3 / 摘要 --- p.5 / Content --- p.6 / List of Abbreviations and symbols --- p.10 / Chapter Chapter 1 --- Introduction --- p.13 / Chapter 1.1 --- The Severity of Influenza A Virus --- p.14 / Chapter 1.2 --- Introduction to Influenza A Virus --- p.15 / Chapter 1.3 --- What is Nucleoprotein? --- p.17 / Chapter 1.4 --- Multifunctional role of Nucleoprotein --- p.19 / Chapter 1.4.1 --- Interaction of Nucleoprotein with Other Viral Components --- p.19 / Chapter 1.4.1 --- Interaction of Nucleoprotein with Cellular Components --- p.22 / Chapter 1.5 --- Aims of study --- p.23 / Chapter Chapter 2 --- Materials and Methods / Chapter 2.1 --- Materials --- p.25 / Chapter 2.1.1 --- Chemical reagents --- p.25 / Chapter 2.1.2 --- Buffers --- p.28 / Chapter 2.1.2.1 --- Preparation of Buffers --- p.28 / Chapter 2.1.2.2 --- Buffer for Common Use --- p.28 / Chapter 2.1.3 --- Plasmids and Strains --- p.31 / Chapter 2.2 --- Methods --- p.32 / Chapter 2.2.1 --- Molecular Cloning --- p.32 / Chapter 2.2.2 --- "Expression of the Recombinant NP-WT/ mutants and importin α1, 3and 5 in E.coli" --- p.46 / Chapter 2.2.3 --- Purification of the NP WT/ variants --- p.50 / Chapter 2.2.4 --- "Purification of importin αl, 3 and 5" --- p.53 / Chapter 2.2.5 --- In vitro interaction study between NP and importin α --- p.56 / Chapter 2.2.6 --- In vivo interaction study between NP and importin α --- p.59 / Chapter 2.2.7 --- In vivo analysis to study NP-NP homo-oligomerization --- p.64 / Chapter 2.2.8 --- In vitro static light scattering analysis to determine NP oligomeric state --- p.68 / Chapter 2.2.9 --- Control experiments to verify NP-polymerases and NP-RNA interaction --- p.69 / Chapter Chapter 3 --- Functional analysis of influenza H5N1 nucleoprotein tail loop for oligomerization and ribonucleoprotein activities / Chapter 3.1 --- Introduction --- p.72 / Chapter 3.2 --- Results --- p.75 / Chapter 3.2.1 --- Tail loop insertion is maintained by intra- and inter-molecular interactions --- p.75 / Chapter 3.2.2 --- NP mutants display defective transcription-replication activity --- p.77 / Chapter 3.2.3 --- Expression and purification of defective NP variants --- p.80 / Chapter 3.2.4 --- Defective NP variants interact with RNA and the polymerase complex --- p.85 / Chapter 3.2.5 --- Defective NP variants possess abnormal oligomeric states in vitro --- p.91 / Chapter 3.2.6 --- NP variants with impaired RNP activity cannot form homo-oligomers in vivo --- p.95 / Chapter 3.3 --- Discussion --- p.98 / Chapter Chapter 4 --- Biophysical characterization of the interaction between influenza nucleoprotein and importin α / Chapter 4.1 --- Introduction --- p.105 / Chapter 4.2 --- Results --- p.108 / Chapter 4.2.1 --- Expression and Purification of NP-WT/ NLS variants and Importin a --- p.108 / Chapter 4.2.2 --- Pull down Assay --- p.113 / Chapter 4.2.3 --- Light scattering analysis (NP-lmportin α5) --- p.114 / Chapter 4.2.4 --- Binding assay of NP NLS variants with importin α5 --- p.115 / Chapter 4.2.5 --- BIAcore 3000 Surface Plasmon Resonance (NP-lmportin α5) --- p.118 / Chapter 4.2.6 --- QRT-PCR (NP-lmportin a5) --- p.123 / Chapter 4.3 --- Discussion --- p.125 / References --- p.134
| Identifer | oai:union.ndltd.org:cuhk.edu.hk/oai:cuhk-dr:cuhk_327131 |
| Date | January 2010 |
| Contributors | Chan, Wai Hon., Chinese University of Hong Kong Graduate School. Division of Life Sciences. |
| Source Sets | The Chinese University of Hong Kong |
| Language | English, Chinese |
| Detected Language | English |
| Type | Text, bibliography |
| Format | print, 141, [1] leaves : ill. ; 30 cm. |
| Rights | Use of this resource is governed by the terms and conditions of the Creative Commons “Attribution-NonCommercial-NoDerivatives 4.0 International” License (http://creativecommons.org/licenses/by-nc-nd/4.0/) |
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