心肌型脂肪酸结合蛋白的原核表达与应用研究

发布时间：2018-07-25 10:07

【摘要】：心肌型脂肪酸结合蛋白(H-FABP)是一种低分子量的蛋白质,在脂肪酸的运输、脂类的代谢、调节细胞增殖与分化等方面具有重要的作用,是检测急性心肌梗塞病(AMI)的有效新型生物学标志物。本文根据已报道的心肌型脂肪酸结合蛋白(H-FABP)基因序列,采用原核表达的方式获得了目标蛋白H-FABP。并通过噬菌体表面展示随机十二肽库对H-FABP特异性亲和配体进行了筛选,经四轮筛选及序列基序分析,我们得到两个ELISA阳性特征序列: W-P-N-H-H-M-L-H-K-R-W-P和W-P-H-Q-K-L-H-L-M-R-H-S。随后采用固相法合成这两个标记上芘(一种荧光性很强的化学物质)的目标肽,经高效液相纯化、冻干及质谱分析确定其分子量后作为荧光探针检测AMI阳性病人血样,结果均呈明显的阳性。本研究为AMI临床检测探讨了一种新型、快捷的方法,具有重要的现实意义。
[Abstract]:Myocardial fatty acid binding protein (H-FABP) is a low molecular weight protein, which plays an important role in the transport of fatty acids, metabolism of lipids and regulation of cell proliferation and differentiation. It is an effective new biological marker for detecting (AMI) in acute myocardial infarction. Based on the reported sequence of myocardial fatty acid binding protein (H-FABP) gene, the target protein H-FABPwas obtained by prokaryotic expression. The specific affinity ligands of H-FABP were screened by phage display random dodecapeptide library. After four rounds of screening and sequence analysis, we obtained two ELISA positive characteristic sequences: W-P-N-H-H-M-L-H-K-R-W-P and W-P-H-Q-K-L-H-L-L-M-R-H-S. The two target peptides labeled with pyrene (a highly fluorescent chemical substance) were synthesized by solid phase method. The molecular weight of the peptide was purified by high performance liquid phase, and its molecular weight was determined by lyophilization and mass spectrometry. The molecular weight of the peptide was determined as a fluorescent probe to detect the blood samples of AMI positive patients. The results were obviously positive. This study is a new and fast method for clinical detection of AMI, which has important practical significance.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2006
【分类号】：R341;Q786

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