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Omics Approach to Understand the Formation of Colony Boundar

發(fā)布時(shí)間:2025-01-20 13:21
  各種各樣的細(xì)菌生活在同一個(gè)微生物群落中,它們需要在有限的空間和資源之中進(jìn)行激烈的競爭或合作。相互合作的細(xì)胞可以產(chǎn)生大量有利于其自身生存的行為,包括營養(yǎng)的獲得、群體運(yùn)動(dòng)、毒力、防御、離子吸收、生物膜形成以及子實(shí)體形成。為了確保這些合作行為發(fā)生在同種細(xì)胞之間,細(xì)菌需要被相鄰的細(xì)胞所識(shí)別。微生物社會(huì)學(xué)直接受到鄰近細(xì)胞的協(xié)同或?qū)剐袨榈挠绊?這些行為基于微生物自我識(shí)別和非自我排除以形成相關(guān)群落的能力。細(xì)菌中細(xì)胞間的親緣識(shí)別通常表現(xiàn)為菌落融合不兼容,直到目前,有一些關(guān)于菌落融合不兼容的研究認(rèn)為,同類菌株的兩個(gè)菌落相遇后可以融合,而兩個(gè)非同類菌株的菌落相遇后會(huì)形成一條肉眼可見的界線。一些細(xì)菌物種的近親,例如奇異變形桿菌、枯草芽孢桿菌和黃色粘球菌,能夠在彼此無法識(shí)別的不同菌株的菌落之間形成可見的界線,這就是菌落融合不兼容現(xiàn)象。黃色粘球菌DK1622是一種革蘭氏陰性土壤細(xì)菌,屬于變形桿菌(Proteobacteria)的δ分枝,該模式菌株的基因組己完全測(cè)序。黃色粘球菌的獨(dú)特特征之一是它具有多細(xì)胞行為且是眾所周知的社會(huì)性細(xì)菌,例如在營養(yǎng)豐富的固體表面上,它具有細(xì)胞的群體運(yùn)動(dòng)能力,而在營養(yǎng)成分稀少的情況下,...

【文章頁數(shù)】:190 頁

【學(xué)位級(jí)別】:博士

【文章目錄】:
摘要
Abstract
Chapter 01 INTRODUCTON
    1. Overview
        1.1 PROTEOMIC
            1.1.1 An overview of proteomic approaches
            1.1.2 Post-Translation Modifications
            1.1.3 Mass Spectrometry-based Proteomics
            1.1.4 Database Searching and Protein Identification
            1.1.5 Application of proteomics (overview)
        1.2 METABOLOMICS
            1.2.1 Overview of metabolomics
            1.2.2 Advantages of metabolomics
            1.2.3 Metabolomics techniques
            1.2.4 Applications of metabolomics
        1.3 MYXOBACTERIA
            1.3.1 Overview of myxobacteria
            1.3.2 Diversity and ecology
            1.3.3 Myxobacterial genome
            1.3.4 Social interactions
            1.3.5 Predation
            1.3.6 Production of natural products
        1.4 COLONY-MERGE INCOMPATIBILITY
            1.4.1 Overview of boundary formation
            1.4.2 Balance in colony boundaries
            1.4.3 Mechanism of colony merging incompatibility
        1.5 AIM AND OBJECTIVES
Chapter 02 MATERIAL AND METHODS
    2.1 Materials and chemicals
        2.1.1 Reagents preparation
        2.1.2 Medium and reagents
        2.1.3 Apparatus and instruments
        2.1.4 Strains and plasmids
        2.1.5 List of primers
    2.2 Experimental methods
        2.2.1 Fluorescence microscopy observation of colony boundaries
        2.2.2 Scanning Electron Microscopy (SEM)
        2.2.3 Sample preparation for LC-MS/MS
            2.2.3.1 Purification of digested proteins and peptides by Zip-Tip
        2.2.4 Nano-LC-LTQ-Orbitrap parameters and data processing
        2.2.5 Sample preparation and extraction of compounds from boundary
        2.2.6 High-resolution HPLC-MS/MS system parameters
        2.2.7 T6SS-knockout sample preparation
    2.3 Data analysis
Chapter 03 RESULTS
    3.1 Role of MXAN-0049 gene in colony boundary formation
    3.2 Colony boundaries between incompatible M. xanthus strains under microscope
    3.3 Proteomic analysis of colony boundaries
        3.3.1 Significantly expressed protein within boundary
        3.3.2 Gene ontology of boundary
        3.3.3 Significance proteins and their co-relation with boundary formation
        3.3.4 Proteomics analysis and KEGG pathways
        3.3.5 Role of Protein Export pathway in boundary
        3.3.6 Interaction of Cold shock proteinswith RNA degmdation pathway
        3.3.7 Co-relationships of boundary formation with T6SS system
        3.3.8 Phenylalanine, tyrosine and tryptophan biosynthesis
    3.4 Identification of chemical compounds by HPLC-MS/MS
        3.4.1 antiSMASH prediction of gene cluster and boundary
        3.4.2 Confirmation of aromatic amino acids production within boundary
        3.4.3 Interaction of groEL1 and groEL2 with △MXAN0049
        3.4.4 Significantly produced unknown compounds within boundary
CHAPTER 04 DISCUSSION
    4.1 Limitations and
    4.2 Future directions
APPENDIX-Ⅰ
    List of Abbreviation
    List of figures
    List of tables
REFERENCES
ACKNOWLEDGEMENT
LIST OF PUBLICATION
學(xué)位論文評(píng)閱及答辯情況表



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