结核分枝杆菌主要分泌蛋白的功能和免疫原性分析.pdfVIP

Huazhong Agricultural University, 2020 Ph.D. Dissertation CONTENT 摘 要 I ABSTRACT III ABBREVIATIONV CHAPTER 1: INTRODUCTION 1 1.1 INTRODUCTION1 1.2 Literature review 2 1.2.1 History2 1.2.2 Transmission and Pathogenesis3 1.2.3 Epidemiology and topographical spread 5 1.2.5 Vaccines for TB 9 1.2.6 Peptide vaccine(s) for TB11 1.2.7 Genomic organization features12 1.2.8 Functional genetics of Mtb 13 Objectives:20 CHAPTER 2: SUBUNIT VACCINE DESIGN AND IN SILICO ANALYSIS21 2.1 MATERIALS AND METHODS21 2.1.1 Collection of protein sequences for vaccine design 21 2.1.2 MHC-I/-II binding and B cell epitopes prediction 21 2.1.3 Multi-epitope subunit vaccine construct21 2.1.4 Prediction of physiochemical parameter, allergenicity, antigenicity analysis 22 2.1.5 Docking of SV with mouse TLR2 receptor 22 2.1.6 Reverse translation, codon optimization and in silico cloning 23 2.2 RESULTS23 2.2.1 Construction of multi-epitope vaccine sequence 23 2.2.2 Physiochemical properties, Allergenicity and antigenicity analysis 26 2.2.3 Molecular docking 26 2.2.4 Reverse translation, codon optimization and in silico cloning 27 CHAPTER 3: IN VITRO WORKING OF SUBUNIT VACCINE28 3.1 MATERIALS AND METHODS 28 3.1.1 In vitro cloning 28 3.1.2 Intracellular SV expression 28 IV Function and Immunogenicity analysis of Mycobacterium tuberculosis main secretory proteins 3.1.3 AAV virus generation 29 3.1.4 DNA and RNA copy number determination 29 3.2 RESULTS 31 3.2.1 Removal of adjuvant and docking of vaccine protein with mouse TLR931 3.2.2 Cloning confirmation 31 3.2.3 Subunit vaccine protein expression 32 3.2.4 Calculated DNA and RNA copy number 32 CHAPTER 4: RV3444C-RV3445C CHARACTERIZATION 34 4.1 MATERIALS AND METHODS 34 4.1.1 Hypot