心肌型脂肪酸結(jié)合蛋白的原核表達(dá)與應(yīng)用研究
發(fā)布時間：2018-07-25 10:07
【摘要】：心肌型脂肪酸結(jié)合蛋白(H-FABP)是一種低分子量的蛋白質(zhì),在脂肪酸的運輸、脂類的代謝、調(diào)節(jié)細(xì)胞增殖與分化等方面具有重要的作用,是檢測急性心肌梗塞病(AMI)的有效新型生物學(xué)標(biāo)志物。 本文根據(jù)已報道的心肌型脂肪酸結(jié)合蛋白(H-FABP)基因序列,采用原核表達(dá)的方式獲得了目標(biāo)蛋白H-FABP。并通過噬菌體表面展示隨機十二肽庫對H-FABP特異性親和配體進(jìn)行了篩選,經(jīng)四輪篩選及序列基序分析,我們得到兩個ELISA陽性特征序列: 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。隨后采用固相法合成這兩個標(biāo)記上芘(一種熒光性很強的化學(xué)物質(zhì))的目標(biāo)肽,經(jīng)高效液相純化、凍干及質(zhì)譜分析確定其分子量后作為熒光探針檢測AMI陽性病人血樣,結(jié)果均呈明顯的陽性。本研究為AMI臨床檢測探討了一種新型、快捷的方法,具有重要的現(xiàn)實意義。
[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.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號】：R341;Q786
本文編號：2143469
[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.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2006
【分類號】：R341;Q786
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，本文編號：2143469
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