本文選題：人白介素-24 + 表達(dá)與純化��； 參考：《吉林大學(xué)》2008年碩士論文

【摘要】： 人白介素-24是一種細(xì)胞因子且屬于白介素-10家族。最初是在人黑色素瘤細(xì)胞中發(fā)現(xiàn)的,該基因在誘導(dǎo)分化的黑色素瘤細(xì)胞中呈高表達(dá),并能促進(jìn)黑色素瘤細(xì)胞分化,因此又稱為黑色素瘤分化相關(guān)基因-7,于2002年被命名為IL-24。 hIL-24全長(zhǎng)為1718bp,其成熟肽編碼一個(gè)158個(gè)氨基酸組成的新蛋白,相對(duì)分子質(zhì)量為18400。IL-24由于具有抑制腫瘤生長(zhǎng)的作用而被認(rèn)識(shí)。針對(duì)多種組織來源的腫瘤,發(fā)現(xiàn)表達(dá)的IL-24可發(fā)揮生長(zhǎng)抑制和誘導(dǎo)凋亡的作用,并且對(duì)正常細(xì)胞沒有毒害作用。hIL-24還具有誘導(dǎo)PBMC分泌IL-6、TNF-α、IFN-r、IL-12等細(xì)胞因子的功能,IL-24基因是一個(gè)既抑制腫瘤細(xì)胞生長(zhǎng),又表達(dá)刺激免疫系統(tǒng)的蛋白的基因。對(duì)該基因的研究具有非常重要的理論和臨床意義。 國(guó)外文獻(xiàn)報(bào)道大多數(shù)是針對(duì)hIL-24腺病毒表達(dá)載體的研究與開發(fā),對(duì)原核表達(dá)研究較少。原核表達(dá)系統(tǒng)具有良好的可操作性,成本低,可獲得大量具有生物學(xué)活性的重組蛋白,并且可以進(jìn)一步開發(fā)為應(yīng)用方便、安全的基因重組藥物。 本實(shí)驗(yàn)利用PCR技術(shù)從含有白介素-24基因組中擴(kuò)增出hIL-24的基因片段(491bp),根據(jù)同尾酶的原理成功地構(gòu)建了hIL-24的原核表達(dá)載體pQE30- hIL-24,采用鎳凝膠親和層析的方法獲得大量純化目的蛋白。
[Abstract]:Human interleukin-24 is a cytokine and belongs to the IL-10 family. Initially found in human melanoma cells, the gene is highly expressed in differentiated melanoma cells and promotes the differentiation of melanoma cells. Therefore, also known as melanoma differentiation related gene -7, it was named IL-24hIL-24 in 2002 as a full-length 1718bp. its mature peptide encodes a new protein composed of 158 amino acids. The relative molecular weight of 1840. 0. IL-24 is known to inhibit tumor growth. It was found that IL-24 could inhibit growth and induce apoptosis in various tumor tissues. Moreover, hIL-24 has no toxic effect on normal cells. It can also induce PBMC to secrete cytokines such as IL-6TNF- 偽 and IFN-r-IL-12. IL-24 gene is a gene that not only inhibits the growth of tumor cells, but also expresses proteins that stimulate the immune system. The study of this gene has very important theoretical and clinical significance. Most of the reports in foreign literature focus on the research and development of hIL-24 adenovirus expression vector, but less on prokaryotic expression. Prokaryotic expression system has good maneuverability, low cost, can obtain a large number of recombinant proteins with biological activities, and can be further developed as a convenient and safe gene recombinant drug. In this experiment, hIL-24 gene fragment (491bp) was amplified from the genome of interleukin-24 by polymerase chain reaction (PCR), and the prokaryotic expression vector pQE30-hIL-24 was successfully constructed according to the principle of isotail enzyme. A large number of purified target proteins were obtained by nickel gel affinity chromatography.
【學(xué)位授予單位】：吉林大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類號(hào)】：R392;Q78

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