أعرض في EDS

Roles of the MSP-1₃₃ in the induction of anti-malaria response.

التفاصيل البيبلوغرافية
العنوان: Roles of the MSP-1₃₃ in the induction of anti-malaria response.
مرشدي الرسالة: Tam, Hou Si., Chinese University of Hong Kong Graduate School. Division of Biochemistry.
سنة النشر: 2007
المجموعة: The Chinese University of Hong Kong
Original Material: cuhk:326094
مصطلحات موضوعية: Malaria, Immunological aspects, Malaria, Falciparum, immunology, Merozoite Surface Protein 1, Peptide Fragments
الوصف: Tam, Hou Si.
33 in title is subscript.
Thesis (M.Phil.)--Chinese University of Hong Kong, 2007.
Includes bibliographical references (leaves 174-187).
in English and Chinese.
THESIS COMMITTEE --- p.i
ACKNOWLEDGEMENTS --- p.ii
p.iii
摘要 --- p.v
TABLE OF CONTENTS --- p.vii
LIST OF FIGURES --- p.xii
LIST OF TABLES --- p.xvii
LIST OF ABBREVIATIONS --- p.xviii
CHAPTER
Chapter 1. --- INTRODUCTION
Chapter 1.1 --- Malaria --- p.1
Chapter 1.2 --- Malaria is a public health problem --- p.1
Chapter 1.3 --- Malarial parasite --- p.3
Chapter 1.4 --- Life cycle of P. falciparum --- p.3
Chapter 1.4.1 --- The pre-erythrocytic stage --- p.3
Chapter 1.4.2 --- The asexual erythrocytic stage --- p.3
Chapter 1.4.3 --- The sexual transmission stage --- p.6
Chapter 1.5 --- Chemoprophylaxis and chemotherapy of malaria --- p.7
Chapter 1.6 --- Drug resistance of malaria parasite --- p.7
Chapter 1.7 --- The progress for malaria vaccine --- p.10
Chapter 1.8 --- Vaccine candidates for asexual erythrocytic stage --- p.11
Chapter 1.9 --- Merozoite Surface Protein-1 (MSP-1) --- p.13
Chapter 1.9.1 --- Structure of MSP-1 --- p.13
Chapter 1.9.2 --- The processing of MSP-1 --- p.17
Chapter 1.9.3 --- MSP-1 as a blood-stage vaccine --- p.19
Chapter 1.9.4 --- The vaccine potency of MSP-133 --- p.23
Chapter 1.10 --- Merits of E. coli expression system --- p.25
Chapter 1.11 --- Aim of study --- p.26
Chapter 2. --- MATERIALS AND METHODS
Chapter 2.1 --- Materials --- p.30
Chapter 2.2 --- Methods --- p.39
Chapter 3. --- EXPRESSION AND PURIFICATION OF RECOMBINANT MSP-l33kv+19 PROTEIN
Chapter 3.1 --- Introduction --- p.63
Chapter 3.2 --- Results
Chapter 3.2.1 --- Construction of pET32a/MSP-l33kv+19 expression vector --- p.64
Chapter 3.2.2 --- SDS-PAGE analysis of the expressed protein --- p.74
Chapter 3.2.3 --- Western blot analysis of the expressed protein --- p.78
Chapter 3.2.4 --- Modification of the expression conditions --- p.78
Chapter 3.2.5 --- Protein purification by IMAC --- p.82
Chapter 3.2.6 --- Cleavage of fusion partner from the rMSP-133kv+19 protein --- p.82
Chapter 3.2.7 --- Verification of non-fused recombinant MSPl33kv+19 protein by N-terminal amino acid sequencing --- p.86
Chapter 3.2.8 --- Separation of target protein from the fusion mixture by IMAC --- p.86
Chapter 3.2.9 --- Separation of digestion product by Size Exclusion Chromatography --- p.89
Chapter 3.2.10 --- Conformational test of the purified protein --- p.89
Chapter 3.2.11 --- Separation of target protein from contaminants by Anion-Exchange Chromatography --- p.92
Chapter 3.2.12 --- Separation of target protein from contaminants by Immuno-Affinity Chromatography --- p.95
Chapter 3.3 --- Conclusion --- p.95
Chapter 4. --- IMMUNOLOGICAL CHARACTERIZATION OF BACTERIAL EXPRESSED rMSP-l33kv+19
Chapter 4.1 --- Introduction --- p.97
Chapter 4.2 --- Results
Chapter 4.2.1 --- Immunogenicity of recombinant NfMSP-133kV+19 protein --- p.98
Chapter 4.2.2 --- Specificity of anti-NfMSP-133kv+19 sera to MSP-l33kv. MSP-l33 and MSP-l19 --- p.98
Chapter 4.2.3 --- Cross reactivity of anti-MSP-133kv+19 and anti-BVp42 serum --- p.103
Chapter 4.2.4 --- Competitive ELISA --- p.103
Chapter 4.2.5 --- Test for the presence of inhibitory B-cell epitopes on rMSP-l33kv+19 --- p.111
Chapter 4.2.6 --- In vitro parasitic growth inhibition assay --- p.113
Chapter 4.3 --- Conclusion --- p.115
Chapter 5. --- EXPRESSION AND PURIFICATION OF RECOMBINANT MSP-l33kc+19 PROTEIN
Chapter 5.1 --- Introduction --- p.116
Chapter 5.2 --- Results
Chapter 5.2.1 --- Construction of pET32a/MSP-133kv+19 expression vector --- p.117
Chapter 5.2.2 --- Expression of recombinant MSP-133kc+19 protien (rMSP-133kc+19) --- p.124
Chapter 5.2.3 --- Purification of rMSP-l33kc+19 by IMAC --- p.127
Chapter 5.2.4 --- Cleavage of fusion partner from target protein --- p.127
Chapter 5.2.5 --- Construction of pRSETA/MSP-l3X33kc+19 expression vector --- p.135
Chapter 5.2.6 --- SDS-PAGE analysis of the protein expression --- p.146
Chapter 5.3 --- Conclusion --- p.153
Chapter 6. --- DISCUSSION
Chapter 6.1 --- Expression of rMSP-l33kv+19 --- p.154
Chapter 6.2 --- Purification of rMSP-l3.3kv+19 --- p.156
Chapter 6.3 --- Conformational test of rMSP-133kv+19 --- p.157
Chapter 6.4 --- Biological and immunological activity of NfMSP-133kv+19 --- p.158
Chapter 6.5 --- Expression of rMSP-133kc+19 --- p.166
Chapter 6.6 --- Future prospects --- p.167
REFERENCES --- p.174
APPENDICES
Chapter 1. --- HiTrap NHS-activated HP for ligand coupling procedure --- p.188
Chapter 2. --- Reuse of Ni+-NTA Resin procedure --- p.190
Chapter 3. --- Sequence alignment of MSP-133 (MAD20 & Welcome/Kl alleles) --- p.191
Chapter 4. --- Nucleotide sequence and amino acid sequence of P. falciparum MSP-l33kv+19 --- p.192
Chapter 5. --- Nucleotide sequence and amino acid sequence of P. falciparum MSP-l33kc+19 --- p.193
Chapter 6. --- "Nucleotide sequence and amino acid sequence of P. falciparum MSP-142 (3D7 isolate, MAD20 allele)" --- p.194
Chapter 7. --- Amino acid sequence of Plasmodium falciparum MSP-l42 --- p.195
Original Identifier: oai:cuhk-dr:cuhk_326094
نوع الوثيقة: Text
bibliography
وصف الملف: print; xx, 195 leaves : ill., maps ; 30 cm.
اللغة: Chinese
English
الإتاحة: http://library.cuhk.edu.hk/record=b5893276Test
http://repository.lib.cuhk.edu.hk/en/item/cuhk-326094Test
حقوق: 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/4.0Test/)
رقم الانضمام: edsndl.cuhk.edu.hk.oai.cuhk.dr.cuhk.326094
قاعدة البيانات: Networked Digital Library of Theses & Dissertations
الوصف
الوصف غير متاح.