本研究克隆了中國林蛙（Rana Chensinensis） Dmrtl （double-sex and mab-3related transcription factor1）基因,并檢測了該基因在中國林蛙成體不同組織的表達以及在精巢中的定位。然后,將林蛙暴露于不同濃度的辛基酚（octylphenol, OP,10-8,10-7,10-6,10-5mol/L）后,檢測精巢中Dmrtl mRNA的表達變化,探討OP對Dmrt1基因表達的影響,分析Dmrt1可能的生理功能。采用反轉(zhuǎn)錄聚合酶鏈式反應（RT-PCR）和cDNA拼接技術(shù),克隆中國林蛙Dmrt1基因,對其所編碼的氨基酸序列信息,進化關(guān)系,蛋白質(zhì)理化性質(zhì),二三級結(jié)構(gòu),親水/疏水性,跨膜結(jié)構(gòu),信號肽,細胞定位進行分析。通過RT-PCR分析Dmrt1在成體精巢、卵巢、肝臟、腎臟和腦組織中的分布；對幼體性腺中腎復合體（GMC）中Dmrt1的表達變化也用RT-PCR進行分析；雌雄基因組中Dmrt1水平的差異用PCR技術(shù)進行簡單評估。用熒光實時定量PCR檢測不同OP濃度和不同暴露時間對Dmrt1mRNA表達的影響；用原位雜交技術(shù)檢測Dmrt1m... 

【文章來源】：陜西師范大學陜西省 211工程院校 教育部直屬院校

【文章頁數(shù)】：71 頁

【學位級別】：碩士

【文章目錄】：
摘要
Abstract
Unit 1 Review on estrogen effects of 4-tert-octylphenol (OP)on reproductive and development in animal
    1. The distribution of OP and its effect of estrogen
    2. The effects of OP on animal reproductive and development
    3. The mechanism of endocrine activity interfered by OP
        3.1 OP paly roles in combination with estrogen receptor(ER)
        3.2 OP paly roles by interfering with the synthesis and metabolic pathway of steroid hormones
        3.3 Other possible ways
    4. The problems and prospect
Unit 2 Clonging and expression pattern of Double-sex and mab-3 relatedtranscription factor 1(Dmrt1)in frog Rana Chensinensis
    1. Introduction
    2. Materials and methods
        2.1 Materials
        2.2 Method
    3. Results and analysis
        3.1 The results of total RNA extraction
        3.2 Cloning and structure analysis of a full-length Dmrt1
        3.3 Dmrt1 expression in different tissues
        3.4 Expression analysis of Dmrt1 during R.chensinensis gonad differentiation and development
        3.5 Dmrt1 gene cloning from genomic DNA
    4. Discussion
    5. Conclusion
Unit 3 Effects of 4-tert-octylphenol on Dmrtl mRNAexpression in testes of frog Rana Chensmensis
    1. Introduction
    2. Materials and methods
        2.1 Materials
        2.2 OP exposure methods and sample collection
        2.3 In situ hybridization
        2.4 Real time fluorescent quantitative RT-PCR
        2.5 Photography and Statistical analysis
    3. Results
        3.1 The results of in situ hybridization of Dmrt1 mRNA
        3.2 Effects of OP on Dmrt1 mRNA expression
    4. Discussion
    5. Conclusion
Summarize
References
致謝
攻讀碩士學位期間的研究成果


【參考文獻】：
期刊論文
[1]兩棲動物性別決定相關(guān)基因的研究進展[J]. 劉佳,李忻怡,張育輝.  動物學雜志. 2011(06)
[2]壬基酚對中國林蛙精巢中StAR表達的影響[J]. 李忻怡,王慧,張育輝.  環(huán)境科學研究. 2011(05)
[3]Distribution patterns of octylphenol and nonylphenol in the aquatic system at Mai Po Marshes Nature Reserve,a subtropical estuarine wetland in Hong Kong[J]. LI Xiang-li1,2,LUAN Tian-gang2,LIANG Yan1,,WONG Ming-hung1,LAN Chong-yu2,1.Croucher Institute for Environmental Sciences and Department of Biology,Hong Kong Baptist University,Hong Kong,China.2.State Key Laboratory of Biocontrol,School of life Sciences,Sun Yat-Sen(Zhongshan)University,Guangzhou 510275,China..  Journal of Environmental Sciences. 2007(06)



本文編號：3515582