蘋果酸脫氫酶（MDH,EC 1.1.1.37）屬于A類脫氫酶,它可以組成具有高度保守性的NAD（P）⁺依賴性的基因家族,可催化草酰乙酸鹽和蘋果酸鹽的可逆轉(zhuǎn)換。而蘋果酸是植物細(xì)胞中重要的中間代謝物,在乙醛酸循環(huán)、三羧酸循環(huán)、葡萄糖合成、氨基酸合成以及氧化還原穩(wěn)定性等細(xì)胞代謝途徑中具有多種生物學(xué)功能。目前,關(guān)于棉花MDH基因家族少有研究。本研究對棉花MDH基因家族進(jìn)行了全基因組鑒定及比較分析,闡明了該基因家族在棉花基因組內(nèi)的組成以及在棉纖維不同生長時期的表達(dá)特征,為該基因家族在棉纖維細(xì)胞發(fā)育過程中的功能研究奠定了基礎(chǔ)。在雷蒙德氏棉、亞洲棉和陸地棉基因組中共鑒定到51個MDH基因。多重序列比對結(jié)果顯示棉花MDH基因可以分為五個亞組:線粒體MDH,過氧化物酶體MDH,質(zhì)體MDH,葉綠體MDH和細(xì)胞質(zhì)MDH。相同亞組內(nèi)的MDH基因在基因結(jié)構(gòu)、氨基酸序列以及功能結(jié)構(gòu)域的保守基序方面具有相似的特征。染色體定位分析暗示基因片段復(fù)制事件在棉花MDH基因家族擴(kuò)張過程中發(fā)揮了主要作用,而純化選擇在棉花MDH基因家族隨后的進(jìn)化過程中也起到了重要作用。進(jìn)一步的表達(dá)分析結(jié)果顯示陸地棉MDH基... 

【文章來源】：清華大學(xué)北京市 211工程院校 985工程院校 教育部直屬院校

【文章頁數(shù)】：108 頁

【學(xué)位級別】：博士

【文章目錄】：
摘要
Abstract
Chapter 1 Introduction
    1.1 Malate dehydrogenase gene families in plants
        1.1.1 Plant malate dehydrogenase gene families
        1.1.2 Plant malate dehydrogenase functions
        1.1.3 Research progresses of cotton malate dehydrogenase
    1.2 Cotton fibers
        1.2.1 Cotton fiber development
        1.2.2 Cotton fiber elongation mechanism
        1.2.3 Importance of malate dehydrogenase during fiber elongation
    1.3 Cotton genomes
        1.3.1 Molecular evolution of cotton genome
        1.3.2 Sequence date of cotton genome
    1.4 Research objectives and proposed significance
        1.4.1 Key problems
        1.4.2 Research purposes and contents
        1.4.3 Proposed significance
Chapter 2 Material and methods
    2.1 Biomaterial
    2.2 Bioinformatics analysis
    2.3 RACE PCR
    2.4 Real-time quantitative and semi-quantitative RT-PCR
    2.5 Subcellular localization analysis
    2.6 Analysis of protein expression
    2.7 MDH activity assay and determination of malate content
    2.8 Prokaryotic expression and purification of proteins
Chapter 3 Identification of cotton MDH gene family and their sequence features
    3.1 Introduction
    3.2 Identification of MDH gene family in G. raimondii and G. arboretum genomes
    3.3 Identification of MDH gene family in G. hirsutum genome
    3.4 Sequence characteristics of cotton MDH genes
    3.5 Conserved domain analysis of cotton MDH genes
    3.6 Summary
Chapter 4 Evolutionary relationship and expression profiles of MDH gene family
    4.1 Introduction
    4.2 Phylogenetic relationships of cotton MDH genes
    4.3 Chromosomal location and duplication analysis of cotton MDH genes
    4.4 Expression pattern of MDH genes in different tissues of allotetraploid cotton
    4.5 Putative regulatory cis-elements in MDH gene promoters
    4.6 Summary
Chapter 5 Molecular characterization of GhcMDH1 gene
    5.1 Introduction
    5.2 Molecular cloning of a GhcMDH1 gene
    5.3 GhcMDH1 sequence features and phylogenetic analysis
    5.4 Tissue expression and subcellular localization of GhcMDH1
    5.5 MDH enzymatic activity and malate content in cotton fiber cells
    5.6 Summary
Chapter 6 Discussion and conclusions
    6.1 Discussion
    6.2 Main conclusions
    6.3 Future prospects
References
Acknowledgement
Appendix I
Appendix II
Personal resume and published papers


【參考文獻(xiàn)】：
期刊論文
[1]The Gossypium raimondii Genome,a Huge Leap Forward in Cotton Genomics[J]. Yu-Xian Zhu,Fu-Guang Li.  Journal of Integrative Plant Biology. 2013(07)
[2]Gene clone, expression and enzyme activity assay of a cytosolic malate dehydrogenase from apple fruits[J]. Yuxin YAO, Yujin HAO, Ming LI, Mingli PANG, Zhi LIU, Heng ZHAI State Key Laboratory of Crop Biology, College of Horticultural Science and Engineering, Shandong Agricultural University, Tai’an 271018, China.  Frontiers of Agriculture in China. 2008 (03)
[3]GSDS:基因結(jié)構(gòu)顯示系統(tǒng)[J]. 郭安源,朱其慧,陳新,羅靜初.  遺傳. 2007(08)
[4]KaKsCalculator:Calculating Ka and Ks Through Model Selection and Model Averaging[J]. Gane Ka-Shu Wong.  Genomics Proteomics & Bioinformatics. 2006(04)



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