药学专业毕业论文+任务书+开题报告+文献综述+外文翻译26.docVIP

本科毕业论文（设计）手册 ( 届) 题 目： 重组融合蛋白TAT-HA-aFGF27-154的1.材料………………………………………………………………………………7 1.1 融合蛋白和细胞…………………………………………………………………7 1.2主要试剂…………………………………………………………………………7 1.2.1细胞培养相关试剂………………………………………………………7 1.2.2 主要仪器设备………………………………………………………………7 2．实验步骤…………………………………………………………………………8 2.1 融合蛋白的除盐与浓缩………………………………………………………8 2.2 融合蛋白的冻干………………………………………………………………8 2.3 3T3细胞的培养………………………………………………………………8 2.4 MTT测活……………………………………………………………………8 2.5 统计学分析…………………………………………………………………9 3．实验结果…………………………………………………………………………9 3.1 3T3细胞的体外培养…………………………………………………………9 3.2 融合蛋白的MTT测活………………………………………………………9 4．讨论………………………………………………………………………………14 参考文献……………………………………………………………………………16 致谢……………………………………………………………………………………18 重组融合蛋白TAT-HA-aFGF27-154的生物学活性检测 摘 要：目的：对早期表达的融合蛋白TAT-HA-aFGF27-154进行细胞活性检测，找出最佳的药物浓度，为进一步的生物学应用提供理论基础。方法：纯化得到的融合蛋白TAT-HA-aFGF27-154通过超滤除盐并浓缩，冻干后于-80℃保存备用。体外培养3T3细胞，MTT法检测融合蛋白TAT-HA-aFGF27-154的活性。结果：表达、纯化得到的融合蛋白TAT-HA-aFGF27-154去除了野生型aFGF对NIH/3T3细胞的分裂活性，在低浓度条件条件下，对NIH 3T3细胞没有明显的促增殖作用。在外源加入肝素的刺激下，相同浓度的融合蛋白促NIH/3T3细胞分裂的活性增加，其促增殖活性呈浓度依赖性。结论：本研究成功建立融合蛋白TAT-HA-aFGF27-154的体外测活方法，为利用TAT作为药物载体，以及非促分裂型aFGF应用于临床提供有效方法。MTT法细胞测活Recombination fusion protein TAT - HA - aFGF27-154 biological activities detection Abstract：objectiveTAT - HA - aFGF27-154 on NIH/3T3 and to find out the best drug concentration for further in vivo experiment. Methods: the fusion protein TAT - HA - aFGF27-154 was purified through ultrafiltration for demineralization and enrichment, and preserved at -80℃ after lyophilizatio. 3T3 cells were cultured in vitro, and MTT assay was used to determine the mitogenic effect of TAT - HA - aFGF27-154. Results: According to the result of the MTT assay on 3T3 cells, the mitogenic effect of wildtype aFGF was removed in TAT - HA - aFGF27-154 removed, in the condition of low concentration. However, with the stimulation of exogenous heparin, the mitogenic effect of TAT - HA - aFGF27-154 restored in a dose dependent way. Conclusion: this study successfully set up an in vitro biological test meth