轉(zhuǎn)基因（GENETICALLY MODIFIED,GM）作物的分子特征（包括插入位點(diǎn)和旁側(cè)序列等）是GM作物安全評(píng)估和監(jiān)管的基本組成部分,其表征必須以個(gè)案的方式予以進(jìn)行�，F(xiàn)有的各種基于聚合酶鏈反應(yīng)（POLYMERASE CHAIN REACTION,PCR）的GM作物分子特征的表征方法都具有獨(dú)特的優(yōu)缺點(diǎn),任何一個(gè)單獨(dú)的PCR方法幾乎都不可能完全揭示一個(gè)給定GM作物的分子特征,特別是碰到宿主基因組復(fù)雜或有非預(yù)期插入的GM事件。最近開(kāi)發(fā)的新一代測(cè)序技術(shù)（NEW GENERATION SEQUENCING,NGS）為GM作物分子特征的表征提供了一種潛在的替代方案,其分辨率遠(yuǎn)高于基于PCR的方法。NGS已成功應(yīng)用于GM大豆和GM水稻等作物的分子特征表征,但由于玉米基因組的復(fù)雜,NGS在GM玉米分子特征表征方面的應(yīng)用很少。在本研究中,我們將基于PCR和基于NGS的方法結(jié)合起來(lái),試圖揭示我國(guó)自主研發(fā)的兩個(gè)GM玉米事件的分子特征（主要是插入位點(diǎn)和旁側(cè)序列）。這兩個(gè)GM玉米分別是IE09S034和12-5,前者是抗蟲(chóng)GM玉米,后者是耐除草劑和抗蟲(chóng)雙效GM玉米。所用的PCR方法包括熱不對(duì)稱(chēng)交錯(cuò)PCR（T... 

【文章來(lái)源】：上海交通大學(xué)上海市 211工程院校 985工程院校 教育部直屬院校

【文章頁(yè)數(shù)】：110 頁(yè)

【學(xué)位級(jí)別】：碩士

【文章目錄】：
摘要
Abstract
List of Abbreviations
Chapter 1 Introduction
    1.1 Worldwide Status and Division of Commercial Transgenic Crops along with its Economic Background
    1.2 Agricultural Biotechnology in China
        1.2.1 Overview of Agricultural Biotechnology in China
        1.2.2 Challenges Towards Commercialization of GM Crops in China
        1.2.3 Risk Assessment of Biotech Crops in China
        1.2.4 Labelling Policies of Biotech Crops in China
        1.2.5 GM Products in China and Scientist Perspective
    1.3 Past and Present of the Detection Methods for GM Crops and Food
        1.3.1 Introduction to Molecular Characterization of GM
        1.3.2 Methods for the Molecular Characterization of Gmo’s
        1.3.3 DNA based Methods for Identification of Unknown Flanking Sequence and Insertion Site
        1.3.4 Protein Based Detection Methods
        1.3.5 PCR Based Molecular Characterization Methods
        1.3.6 Next Generation Sequencing
    1.4 Objectives
Chapter 2 Materials and Methods
    2.1 Materials
        2.1.1 IE09S034
        2.1.2 12-5
    2.2 Methods
        2.2.1 Dna Extraction
        2.2.2 Reagents and Equipment
        2.2.3 Tail-PCR
        2.2.4 Inverse Polymerase Chain Reaction
        2.2.5 Digital PCR
        2.2.6 Southern Blot
        2.2.7 Next Generations Sequencing
Chapter 3 Results
    3.1 Result of IE09S034 event
        3.1.1 Validation of full-length of the insertion
        3.1.2 Southern blot results
        3.1.3 Digital pcr results
        3.1.4 Insertion site and flanking sequence identified by tail-pcr
        3.1.5 Insertion site and flanking sequence identified by i-pcr
        3.1.6 Insertion site and flanking sequence identified by NGS
    3.2 Results of12-5 event
        3.2.1 VALIDATION OF CRY1AB/CRY2AJ AND G10-EVO-EPSPS IN 12-5
        3.2.2 Validation of sequence between CAMV35S terminator and CAMV35S/Ubiquitin mixed promotor
        3.2.3 Validation of sequence between CAMV35S/Ubiquitin mixed promotor and PEPC
        3.2.4 Validation of sequence between PEPC and CRY1AB/CRY2AJ
        3.2.5 Validation of sequence between CRY1AB/CRY2AJ and maize Ubiquitin promotor pZmUbi-1
        3.2.6 Southern blot results
        3.2.7 Digital pcr results
        3.2.8 Insertion site and flanking sequence identified by tail-pcr
        3.2.9 Insertion site and flanking sequence identified by I-pcr
        3.2.10 Insertion site and flanking sequence identified by NGS
Chapter 4 Discussion and Conclusion
Acknowledgements
References
Publications
附錄


【參考文獻(xiàn)】：
期刊論文
[1]Global challenges of food safety for China[J]. Joseph J.JEN.  Frontiers of Agricultural Science and Engineering. 2018(03)



本文編號(hào)：3559908