利用基因组生物信息学预测和蛋白组学对牛支原体分泌蛋白组的研究及分泌性抗原MbovP0581的验证.pdfVIP

Study on secretome of Mycoplasma bovis with genomic bioinformatics prediction and proteome approach and confirmation of secreted antigenic MbovP0581 Table of Contents Table of Contents I 摘要 IV AbstractV Abbreviations VII 1. Introduction 1 1.1 Background of this research 1 1.1.1 Bovine Mycoplasmosis in cattle population of China 2 1.1.2 Prevention and control strategies of M. bovis 3 1.1.3 Role of genomics and proteomics in the study of bacterial cell secretome 4 1.1.4 Statement of problems 6 1.2 Review of Literature 6 1.2.1 Biological characteristics of Mycoplasmas 6 1.2.2 Pathogenesis of Mycoplasma bovis 11 1.2.3 Diagnosis 15 1.2.4 Vaccination 16 1.2.5 Bacterial cell secretome 18 1.2.6 Secretome of Mycoplasmas 27 1.2.7 Methodology to study secretome of Mycoplasmas 30 2. Aims and objectives 31 3. Identification of antigenic secreted proteins of Mycoplasma bovis with genome and proteome analyses 32 3.1 Introduction 32 3.2 Materials and methods 33 3.2.1 Ethical statement 33 I Huazhong Agricultural University, Ph. D. Dissertation, 2020 3.2.2 Strain and culture conditions 33 3.2.3 Extraction of secretome 33 3.2.4 Protocol for using 2-D clean-up kit 34 3.2.5 2-Dimensional Gel Electrophoresis 35 3.2.6 Matrix-assisted laser desorption/ionization time of flight (MALDI-TOF) mass spectrometry (MS) 35 3.2.7 Bioinformatics analysis 36 3.2.8 Immunoinformatics analysis of secreted proteins 36 3.2.9 Cloning and expression of selected secreted proteins 37 3.2.10 Transformation of ligated sample into BL21 38 3.2.11 Checking protein expression 38