基因表達(dá)的轉(zhuǎn)錄后調(diào)控的一個主要機(jī)制是通過microRNA （miRNA）實(shí)現(xiàn)的。單個的miRNA可以在多種生物學(xué)過程中發(fā)揮關(guān)鍵作用,如細(xì)胞增殖和分化、凋亡和癌變。然而,由哪些特定miRNA可能調(diào)節(jié)羽毛色素沉著尤其是黑色素生成的機(jī)制仍然在很大程度上難以捉摸。因此,本研究旨在比較研究鴨黑色背羽和黑白色背羽二者黑色羽球的miRNA表達(dá)與鴨白色背羽和黑白色背羽二者白色羽球的miRNA表達(dá),這些鴨分別是Cui Hei鴨、改鴨、純系連城鴨和一個改鴨-連城鴨的F2群體。我們運(yùn)用了Solexa測序、生物信息學(xué)和實(shí)驗(yàn)方法初步篩選了與鴨子羽毛色素沉著相關(guān)的miRNA。主要結(jié)果如下：（1）利用MiRDeep2軟件鑒定了129個miRNA,其中121個與已知的雞的miRNA相近,8個為新的miRNA。另外,使用DEGseq軟件還在這兩種組織中鑒定出了五個顯著差異表達(dá)的miRNA,即miR-204-5p、miR-204-3p、miR-144-3p、 miR-2188-3p和miR-130a-5p。值得注意的是,miR-204最主要在黑色羽球中表達(dá)。（2）為了進(jìn)一步驗(yàn)證測序數(shù)據(jù),完成了10個序列已鑒定的miRNA... 

【文章來源】：華中農(nóng)業(yè)大學(xué)湖北省 211工程院校 教育部直屬院校

【文章頁數(shù)】：80 頁

【學(xué)位級別】：碩士

【文章目錄】：
ABSTRACT
中文摘要
ABBREVIATIONS AND ACRONYMS
1 INTRODUCTION
    1.1 Origin of the research question
    1.2 Melanogenesis
        1.2.1 Definition of Melanogenesis
        1.2.2 Melanin synthesis
    1.3 Research progress on candidate genes associated with black and white plumage pigmentation in ducks
    1.4 MicroRNA
        1.4.1 The Discovery of miRNA Regulation
        1.4.2 MiRNA biogenesis
        1.4.3 MiRNA profiling
        1.4.4 MiRNA identified in ducks
        1.4.5 MicroRNA and pigmentation
    1.5 Development and application of high throughput sequencing technology
        1.5.1 Next Generation Sequencing and miRNA Study
        1.5.2 Analysis tools for NGS data
        1.5.3 miRNA Target Prediction
    1.6 Aim of thesis
    1.7 Significance of the research
2 MATERIAL AND METHODS
    2.1 Materials
        2.1.1 Experimental animals
        2.1.2 Software and hardware
        2.1.3 Major reagents
    2.2 Methods
        2.2.1 Preparation of main reagents
        2.2.2 Sample collection,small library preparation
        2.2.3 Sequencing
        2.2.4 Data analysis
            2.2.4.1 Raw data processing and obtaining clean reads
            2.2.4.2 Identification of conserved and novel miRNAs
            2.2.4.3 Differential expression analysis
        2.2.5 Experimental validation of sequencing data by stem loop qPCR
        2.2.6 Tissue profiling of miR-204-5p
        2.2.7 MiRNA target predictions,pathway analysis and GO analysis
        2.2.8 Gene expression analysis and validation of miRNA and target gene interaction.
            2.2.8.1 qPCR
            2.2.8.2 Dual luciferase reporter assay
            2.2.8.3 Western blotting
3 RESULTS
    3.1 Overview of sequencing results
    3.2 Identification of Conserved and Novel miRNAs
    3.3 The first nucleotide bias of miRNA in duck feather bulbs
    3.4 Identification of miRNAs Isoform
    3.5 Identification of differentially expressed miRNAs
    3.6 Experimental validation of sequenced data
    3.7 Tissue expression pattern of miR-204
    3.8 MicroRNAs target predictions,KEGG pathway and GO analysis
    3.9 Expression analysis of miRNA target genes by qPCR
    3.10 Duck miR-27-3pand MITF gene interaction
        3.10.1 Dual luciferase reporter assay
        3.10.2 Western blotting
4 DISCUSSION
    4.1 MiRNAs identified in black and white feather bulbs library of ducks
    4.2 Identification of differentially expressed miRNAs between the feather bulbs
    4.3 MiRNA target prediction
5 SUMMARY
    5.1 The main findings of this study
    5.2 The innovative points of this research
    5.3 Limitations of the study
    5.4 Further research suggestions
CONCLUSION
REFERENCES
PUBLICATIONS
APPENDIX
ACKNOWLEDGEMENT


【參考文獻(xiàn)】：
期刊論文
[1]Characterization of MicroRNA* Species in Peking Duck Skin[J]. ZHANG Li,XIE Xiu-juan,JIA Shan-gang,XIAO Mei,LIN Shu-dai,AN Li-long,LUO Wen,JIA Xin-zheng,NIE Qing-hua,ZHANG Xi-quan.  Journal of Integrative Agriculture. 2013(09)
[2]Identification of Novel and Differentially Expressed MicroRNAs in the Ovaries of Laying and Non-Laying Ducks[J]. YU De-bing,JIANG Bao-chun,GONG Jing,DONG Fu-lu,LU Ying-lin,YUE Hui-jie,WANG Zheng-chao,DU Wen-xing,GUO An-yuan.  Journal of Integrative Agriculture. 2013(01)



本文編號：3598446