Thesis (M.S.)--University of Michigan, 1994. / Includes bibliographical references.

Thesis (Ph.D. in Pharmacology) -- University of Colorado Denver, 2008. / Typescript. Includes bibliographical references (leaves 148-159). Free to UCD Anschutz Medical Campus. Online version available via ProQuest Digital Dissertations;

Thesis (Ph. D.)--University of Washington, 2006. / Vita. Includes bibliographical references (leaves 79-82).

Diss. (sammanfattning) Umeå : Umeå universitet, 2009. / Härtill 4 uppsatser. Även tryckt utgåva.

O Trypanosoma cruzi expressa um grupo de glicoprotcinas de superfície, denominadas Tc-85, que pertencem à superfumília gêmca das gp85/traus-sialidases. Nosso laboratório clonou e caracterizou um membro da fumília Tc85 (Tc85-11), cuja região carboxila tenninal (clone Tc85-1) adere em laminina e em células de mamífero. Usando peptídeos sintéticos, correspondendo em seqüência à Tc85-1, caracterizou-se o motivo mais conservado da superfamilia gênica das gp85/trans-sialidases (VTVxNVFLYNR), o qual não adere em laminina. Esse motivo foi chamado peptídeo J. Por cromatografia de extratos de membrana de cardiomiócitos em coluna de afmidade contendo peptídeo J, foi isolada uma molécula de 30kDa identificada como sendo a subunidade β3 da Na+, K+ ATPase. A porção extracelular da subunidade β3 da Na+, K+ ATPase foi clonada e a interação in vitro desta proteína com peptídeo J foi observada. Deste modo, é sugerido aqui que a subunidade β3 da Na+, K+ ATPase pode ter um papel importante na interação do parasita com a célula hospedeira. / Abstract not available.

Chuck Chi-pang. / Thesis (M.Phil.)--Chinese University of Hong Kong, 2006. / Includes bibliographical references (leaves 138-160). / Abstracts in English and Chinese. / Thesis Committee --- p.ii / Abstract --- p.iii / 摘要 --- p.v / Contents --- p.vii / List of Figures --- p.xi / List of Tables --- p.xiii / Abbreviations --- p.xiv / Acknowledgement --- p.xviii / Introduction / Chapter 1. --- Background / Chapter 1.1 --- SARS / Chapter 1.1.1 --- Outbreak and Influence --- p.1 / Chapter 1.1.2 --- Clinical Features --- p.4 / Chapter 1.2 --- SARS-CoV / Chapter 1.2.1 --- Genomic Organization --- p.5 / Chapter 1.2.2 --- Morphology --- p.7 / Chapter 1.2.3 --- Phylogenetic Analysis --- p.9 / Chapter 1.3 --- S Glycoprotein / Chapter 1.3.1 --- Functional Roles --- p.11 / Chapter 1.3.2 --- Structure and Functional Domains --- p.12 / Chapter 1.3.3 --- Interacting Partners --- p.15 / Chapter 1.3.4 --- Viral Entry Mechanism --- p.17 / Chapter 1.4 --- Aim of Study / Chapter 1.4.1 --- Mismatch of SARS-CoV Tissue Tropism and Tissue Distribution of ACE2 --- p.20 / Chapter 1.4.2 --- Presence of Other Interacting Partner(s) --- p.22 / Chapter 1.4.3 --- Significance of the Study Materials and Methods --- p.22 / Chapter 2. --- Plasmid Construction / Chapter 2.1 --- Fragment Design / Chapter 2.1.1 --- Functional Domain Analysis --- p.23 / Chapter 2.1.2 --- Secondary Structure and Burial Region Predictions --- p.24 / Chapter 2.2 --- Vector Amplification / Chapter 2.2.1 --- E. coli Strain DH5a Competent Cell Preparation --- p.30 / Chapter 2.2.2 --- Transformation of E. coli --- p.30 / Chapter 2.2.3 --- Small-scale Vector Amplification --- p.31 / Chapter 2.3 --- Cloning of DNA Fragments into Various Vectors / Chapter 2.3.1 --- Primer Design --- p.32 / Chapter 2.3.2 --- DNA Amplification --- p.35 / Chapter 2.3.3 --- DNA Purification --- p.35 / Chapter 2.3.4 --- "Restriction Enzyme Digestion, Ligation and Transformation" --- p.36 / Chapter 2.3.5 --- Colony PCR --- p.37 / Chapter 2.4 --- DNA Sequence Analysis / Chapter 2.4.1 --- Primer Design --- p.35 / Chapter 2.4.2 --- DNA Amplification and Purification for DNA Sequence Analysis --- p.39 / Chapter 2.4.3 --- Sequence Detection and Result Analysis --- p.40 / Chapter 3. --- "Protein Expression, Purification and Analysis" / Chapter 3.1 --- Protein Expression in E. coli / Chapter 3.1.1 --- Molecular Weight and pI Predictions --- p.41 / Chapter 3.1.2 --- Glycerol Stock Preparation --- p.41 / Chapter 3.1.3 --- Protein Expression Induction --- p.41 / Chapter 3.1.4 --- Protein Extraction --- p.42 / Chapter 3.1.5 --- Affinity Chromatography --- p.42 / Chapter 3.1.6 --- Removal of GroEL --- p.43 / Chapter 3.1.7 --- Protein Solubilization and Refolding --- p.44 / Chapter 3.2 --- Protein Expression in P. pastoris / Chapter 3.2.1 --- Large-scale Plasmid Amplification --- p.46 / Chapter 3.2.2 --- Restriction Enzyme Digestion and Ethanol Precipitation --- p.47 / Chapter 3.2.3 --- Preparation of KM71H Competent Cells --- p.47 / Chapter 3.2.4 --- Electroporation --- p.48 / Chapter 3.2.5 --- Colony PCR --- p.48 / Chapter 3.2.6 --- Protein Expression Induction and Time Course Study --- p.49 / Chapter 3.2.7 --- Deglycosylation --- p.49 / Chapter 3.3 --- Protein Analysis / Chapter 3.3.1 --- Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis --- p.50 / Chapter 3.3.2 --- Western Blotting --- p.50 / Chapter 3.3.3 --- Mass Spectrometry --- p.51 / Chapter 3.3.4 --- N-terminal Sequencing --- p.52 / Chapter 3.3.5 --- Size Exclusion Chromatography --- p.52 / Chapter 4. --- Identification of Interacting Partner(s) / Chapter 4.1 --- VeroE6 Preparation / Chapter 4.1.1 --- Cell Culture --- p.53 / Chapter 4.1.2 --- Protein Extraction and Western Blotting --- p.53 / Chapter 4.2 --- Pull-down Assay --- p.54 / Chapter 4.3 --- Two-dimensional Gel Electrophores --- p.is / Chapter 4.3.1 --- Isoelectric Focusing --- p.56 / Chapter 4.3.2 --- Sodium Dodecyl Sulfate-Polyacrylamide Gel Electrophoresis --- p.56 / Chapter 4.3.3 --- Silver Staining --- p.57 / Chapter 4.4 --- Mass Spectrometry / Chapter 4.4.1 --- Destaining --- p.58 / Chapter 4.4.2 --- In-gel Digestion --- p.58 / Chapter 4.4.3 --- Desalting by Zip-tip --- p.59 / Chapter 4.4.4 --- Loading Sample --- p.59 / Chapter 4.4.5 --- Peptide Mass Detection and Data Analysis --- p.59 / Results / Chapter 5. --- S Protein Expression / Chapter 5.1 --- Plasmid Construction --- p.61 / Chapter 5.2 --- Molecular Weight and pi Predictions --- p.63 / Chapter 5.3 --- Protein Expression and Optimization in E. coli / Chapter 5.3.1 --- "Comparison of Expression Levels, Solubility and Purities of S Protein Fragments" --- p.64 / Chapter 5.3.2 --- "Alteration of the Solubility in Various Cell Strains, Expression Conditions and Lysis Buffers" --- p.68 / Chapter 5.3.3 --- Identification and Remove of the non-target proteins --- p.72 / Chapter 5.3.4 --- Unfolding and Refolding --- p.79 / Chapter 5.4 --- Protein Expression and Optimization in P. pastoris / Chapter 5.4.1 --- "Expression Levels, Solubility and Purities of Various S Protein Fragments" --- p.85 / Chapter 5.4.2 --- Characterization of De-N-glycosylated Recombinant Proteins --- p.89 / Chapter 6. --- Identification of Interacting partners / Chapter 6.1 --- Practicability of Pull-down Assay / Chapter 6.1.1 --- ACE2 Extraction --- p.95 / Chapter 6.1.2 --- Pull-down of ACE2 by the P. pastoris-expressed recombinant RBD --- p.96 / Chapter 6.2 --- Pull-down Assay and Two-dimensional Gel Electrophoresis --- p.97 / Chapter 6.3 --- Identification of Putative Interacting Partners by MALDI-TOF-TOF --- p.107 / Chapter 7. --- Discussion / Chapter 7.1 --- S Protein Expression in E. coli / Chapter 7.1.1 --- Improving Recombinant Protein Expression Level and Solubility --- p.114 / Chapter 7.1.2 --- S Recombinant Protein Bound by GroEL --- p.117 / Chapter 7.2 --- S Protein Expression in P. pastoris / Chapter 7.2.1 --- Advantages of Using P. pastoris --- p.119 / Chapter 7.2.2 --- Variation of S Fragment Expression Levels --- p.120 / Chapter 7.2.3 --- Sizes of S Protein Fragments --- p.123 / Chapter 7.3 --- Identification of Interacting Partners / Chapter 7.3.1 --- Relationship between S Protein and Putative Interacting Partners --- p.124 / Chapter 7.3.2 --- Failure of Finding ACE2 --- p.125 / Chapter 7.3.2 --- Difficulty in the Identification of Protein Spots --- p.126 / Chapter 7.4 --- Conclusion --- p.131 / Chapter 7.5 --- Future Perspective --- p.132 / Chapter 8. --- Appendix --- p.133 / Chapter 9. --- References --- p.138

Thesis (Ph. D.)--University of Washington, 2003. / Vita. Includes bibliographical references (leaves 98-113).

Diss. (sammanfattning) Umeå : Umeå universitet, 2006. / Härtill 5 uppsatser.

Thesis (Ph. D.)--University of Washington, 2006. / Vita. Includes bibliographical references (leaves 88-101).

O Trypanosoma cruzi expressa um grupo de glicoprotcinas de superfície, denominadas Tc-85, que pertencem à superfumília gêmca das gp85/traus-sialidases. Nosso laboratório clonou e caracterizou um membro da fumília Tc85 (Tc85-11), cuja região carboxila tenninal (clone Tc85-1) adere em laminina e em células de mamífero. Usando peptídeos sintéticos, correspondendo em seqüência à Tc85-1, caracterizou-se o motivo mais conservado da superfamilia gênica das gp85/trans-sialidases (VTVxNVFLYNR), o qual não adere em laminina. Esse motivo foi chamado peptídeo J. Por cromatografia de extratos de membrana de cardiomiócitos em coluna de afmidade contendo peptídeo J, foi isolada uma molécula de 30kDa identificada como sendo a subunidade β3 da Na+, K+ ATPase. A porção extracelular da subunidade β3 da Na+, K+ ATPase foi clonada e a interação in vitro desta proteína com peptídeo J foi observada. Deste modo, é sugerido aqui que a subunidade β3 da Na+, K+ ATPase pode ter um papel importante na interação do parasita com a célula hospedeira. / Abstract not available.