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将硒代半胱氨酸传送进入细胞的一种基于蛋白衣壳的传递系统

摘要第4-8页
ABSTRACT第8-9页
Chapter 1 Introduction第13-29页
    1.1 Nanoparticles in drug delivery第13-15页
    1.2 Virus protein capsid第15-16页
    1.3 Non-Viral protein capsid第16-18页
    1.4 Aa LS protein capsid第18-20页
    1.5 Aa LS-IC第20-21页
    1.6 Selenocystine第21-23页
    1.7 Cell internalization and targeting peptide第23-25页
    1.8 Aim of this thesis第25-29页
Chapter 2 Materials and Methods第29-67页
    2.1 Cells and plasmids第29页
    2.2 Reagents第29-31页
    2.3 Instruments第31-32页
    2.4 Protein production and purification第32-40页
        2.4.1 Materials第32-33页
        2.4.2 Solution preparation第33-35页
        2.4.3 Methods第35-40页
    2.5 Loading selenocystine into Aa LS-IC protein capsid第40-42页
        2.5.1 Materials第40页
        2.5.2 Solution preparation第40页
        2.5.3 Methods第40-42页
    2.6 Determination the reaction yield for Aa LS-IC and selenocystine reaction第42-45页
        2.6.1 Materials第42-43页
        2.6.2 Solution preparation第43页
        2.6.3 Methods第43-45页
    2.7 Structural comparison of Aa LS variants and Sec conjugate第45-48页
        2.7.1 Materials第45页
        2.7.2 Solution preparation第45-46页
        2.7.3 Methods第46-48页
    2.8 Release of selenocysteine第48-53页
        2.8.1 Materials第48页
        2.8.2 Solution preparation第48-49页
        2.8.3 Methods第49-53页
    2.9 Cytotoxicity study第53-57页
        2.9.1 Materials第53-54页
        2.9.2 Solution preparation第54页
        2.9.3 Methods第54-57页
    2.10 Cell uptake study第57-58页
        2.10.1 Materials第57页
        2.10.2 Solution preparation第57页
        2.10.3 Methods第57-58页
    2.11 Synthesis of N-hydroxysuccinimide ester of 7-amino4methyl-coumarin3acetic acid (NHS-AMCA)第58-59页
        2.11.1 Materials第58页
        2.11.2 Methods第58-59页
    2.12 Synthesis of the conjugate between selenocystine and N-hydroxysuccinimideester of 7-amino4methyl-coumarin3acetic acid (NHS-AMCA)第59-67页
        2.12.1 Materials第59页
        2.12.2 Methods第59-67页
Chapter 3 Results第67-101页
    3.1 Loading of selenocystine第67-72页
        3.1.1 Determination of loading yield第67-71页
        3.1.2 Characterization of Aa LS-IC-Sec第71-72页
    3.2 Characterization of Aa LS variants第72-76页
        3.2.1 Structural analysis of Aa LS-EC and Aa LS-EC-Sec第73页
        3.2.2 Structural analysis of Aa LS-IC-pent and Aa LS-IC-pent-Sec第73-76页
    3.3 Release of encapsulated Sec第76-80页
        3.3.1 GSH release of encapsulated Sec第76页
        3.3.2 DTT release of encapsulated Sec第76页
        3.3.3 Negative control for releasing study第76页
        3.3.4 Compare release yield with loading yield第76-80页
    3.4 Cytotoxicity第80-83页
    3.5 Cell uptake第83-84页
    3.6 Synthesis of N-hydroxysuccinimide ester of 7-amino4methyl-coumarin3acetic acid (NHS-AMCA)第84-85页
    3.7 Synthesis of the conjugate between selenocystine and N-hydroxysuccinimideester of 7-amino4methyl-coumarin3acetic acid (NHS-AMCA)第85-101页
        3.7.1 Reaction attempt with wrong selenocystine第85-87页
        3.7.2 Characterization of selenocystine第87-89页
        3.7.3 Reaction attempt for two dye attaching right selenocystine第89-92页
        3.7.4 Control reaction with glycine and cysteine第92-96页
        3.7.5 Reaction attempt for one dye attached selenocystine第96-97页
        3.7.6 Purification for one dye attached selenocystine第97-101页
Chapter 4 Discussion第101-111页
    4.1 Encapsulation and release of Sec第101页
    4.2 Role of capsid architecture and cysteine location第101-102页
    4.3 Correlation between cytotoxicity and cell penetration and location第102-103页
    4.4 Synthesis of the conjugate between selenocystine and N-hydroxysuccinimideester of 7-amino4methyl-coumarin3acetic acid (NHS-AMCA)第103-104页
    4.5 Conclusion & Outlook第104-111页
References第111-121页
Appendix第121-133页
Acknowledgements第133页

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