利用细胞信号转导和通讯机制检测胰高血糖素和皮质醇-分析化学专业论文.docxVIP

山东大学硕士学位论文This 山东大学硕士学位论文 This thesis consists ofthree parts： In chapter onc，an method for determination of glucose was developed based on double enzylneS rcactioll．In this method，we used a solution containing glucose oxidase，horseradish peroxidase and 10-Acetyl·3，7-Dihydroxyphenoxazine．And WC discussed the optimal conditions i．cluding excitation wavelength、concentration of GOD and HRP、the time of incubation In the end of this chapter,we determined the calibralion C1．1rve、位line scope and the nmit ofthe detection ofthe double ellzym嚣 reaction under the optimal conditions． In chapter two，we investigated the cell船1150r of glucagon based on the cell signaling仃锄坞dll商。儿The principle is：After ghcagon combined with special rcccptor Oil the cell surface,the G protein within cells causes the si刚iIlg Wansduction to generate Ouco∞．The generated Ouco∞is released from the cell幽 the solution．The glucose in the solution is then detected using the method described in chapter one．The mount of glucagon can be measured through determining glucose concentration—qince the glucose concentration is higher than the ghcagon concenu．ation,implying amplification of the signal．It was found that the signalⅥfas amplified 50 times． In chapter th∞岛we detected the hydrocortison based on the theory of cell communication and signal transduction．Under the optimal conditions of the expe戚ment,the limit ofdetteclion ofthe ghcagon was 7．0xl旷mol／L and the linear scope was between 1．OxlOS2．3x104 mol／L；the limit of detection of the hydrocorfison was 1．4xlO10 mol／L／ad the linear scope was between 8．Oxlff9 -5．0×10mo]／L．R was found that the signal was amplified 134 times． Keywords：fluorescence sp代臼伪n et|％cell communication,signal transduction 山东大学硬士学位论文符号与缩写 山东大学硬士学位论文 符号与缩写 GOD：葡萄糖氧化酶 HRP：辣根过氧化物酶 ConA：伴刀豆球蛋白 F．RET：共振能量转移 FITC：异硫氰酸荧光素 ADHP：10．乙酰基．3．7．二羟基吩嗪 GGBP：葡萄I／-*-乳糖结合蛋白 cAMP：环磷酸腺苷酸 PKA：蛋白激酶A IP3：三磷酸肌醇 RIAI放射免疫分析 OC：气相色谱 C-C．MS：气质联用技术 原创性声明本人郑重声明：所呈交的学位论文，是本人在导师的指导下，独 原创性声明 本人郑重声明：所呈交的学位论文，是本人在导师的指导下，独 立进行研究