【摘要】： 抗菌肽是生物體產(chǎn)生的一種具有強(qiáng)抗菌作用的陽(yáng)離子多肽,是生物先天免疫的重要組成部分,是機(jī)體理想的第一道防線�？咕木哂蟹肿恿啃�、合成速度快、擴(kuò)散迅速、強(qiáng)堿性、熱穩(wěn)定性高、水溶性好、抗菌機(jī)理獨(dú)特等特性�？咕木哂袕V譜抗菌能力的同時(shí),甚至對(duì)病毒及腫瘤細(xì)胞也能發(fā)揮殺滅作用,更重要的是抗菌肽對(duì)高等動(dòng)物正常細(xì)胞無(wú)明顯的毒害作用。 傳統(tǒng)抗生素耐藥菌株種類(lèi)不斷增多,全球抗生素的濫用加速了這一過(guò)程,抗菌肽的開(kāi)發(fā)將為解決這一棘手難題提供更多的途徑�？咕乃邆涞乃幱脙r(jià)值使其被開(kāi)發(fā)成為新一類(lèi)抗感染藥物成為可能。 本課題通過(guò)人工合成兩種抗菌肽Pexiganan和IB-367的基因,構(gòu)建了表達(dá)載體并篩選了兩株表達(dá)抗菌肽的工程菌株,在大腸桿菌中分別表達(dá)了Pexiganan和IB-367與GST的融合蛋白,摸索出了融合蛋白以可溶形式大量存在于上清中的最佳誘導(dǎo)條件,并通過(guò)親和層析對(duì)表達(dá)載上清中的融合蛋白進(jìn)行純化,純化后的產(chǎn)物經(jīng)過(guò)Factor Xa切割得到兩種目的小肽——Pexiganan和IB-367。抑菌活性檢測(cè)證明Pexiganan和IB-367對(duì)金黃色葡萄球菌及銅綠假單胞菌均有抑制作用。
[Abstract]:Antibacterial peptide is a kind of cationic polypeptide with strong antibacterial effect produced by organisms. It is an important part of biological innate immunity and an ideal first line of defense. Antibacterial peptides are characterized by low molecular weight, rapid synthesis, rapid diffusion, strong alkalinity, high thermal stability, good water solubility and unique antibacterial mechanism. Antibacterial peptides not only have broad spectrum antibacterial ability, but also kill virus and tumor cells. More importantly, antibacterial peptides have no obvious toxic effect on normal cells of higher animals. The abuse of antibiotics in the world has accelerated the development of antibiotic-resistant strains, and the development of antimicrobial peptides will provide more ways to solve this difficult problem. The medicinal value of antibacterial peptides makes it possible to develop a new class of anti-infective drugs. In this paper, two kinds of antibacterial peptides Pexiganan and IB-367 genes were synthesized, the expression vectors were constructed and two engineering strains expressing antibacterial peptides were screened. The fusion proteins of Pexiganan, IB-367 and GST were expressed in E. coli, respectively, and the fusion proteins of Pexiganan, IB-367 and GST were expressed in E. coli. The optimal induction conditions for fusion protein to exist in the supernatant in a large amount in soluble form were found, and the fusion protein expressed in the supernatant was purified by affinity chromatography. The purified products were cleaved by Factor Xa to obtain two small peptides-Pexiganan and IB-367.. The antibacterial activity test showed that both Pexiganan and IB-367 could inhibit Staphylococcus aureus and Pseudomonas aeruginosa.
【學(xué)位授予單位】：天津大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2006
【分類(lèi)號(hào)】：R346

【引證文獻(xiàn)】

相關(guān)博士學(xué)位論文 前1條

1 唐勇;黃鱔腸道cDNA文庫(kù)構(gòu)建、EST分析及抗菌肽基因的原核表達(dá)[D];四川農(nóng)業(yè)大學(xué);2010年

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本文編號(hào)：2465182