人白介素-24原核表达载体的构建、表达和纯化

发布时间：2018-07-01 13:34

本文选题：人白介素-24 + 表达与纯化　；参考：《吉林大学》2008年硕士论文

【摘要】： 人白介素-24是一种细胞因子且属于白介素-10家族。最初是在人黑色素瘤细胞中发现的,该基因在诱导分化的黑色素瘤细胞中呈高表达,并能促进黑色素瘤细胞分化,因此又称为黑色素瘤分化相关基因-7,于2002年被命名为IL-24。 hIL-24全长为1718bp,其成熟肽编码一个158个氨基酸组成的新蛋白,相对分子质量为18400。IL-24由于具有抑制肿瘤生长的作用而被认识。针对多种组织来源的肿瘤,发现表达的IL-24可发挥生长抑制和诱导凋亡的作用,并且对正常细胞没有毒害作用。hIL-24还具有诱导PBMC分泌IL-6、TNF-α、IFN-r、IL-12等细胞因子的功能,IL-24基因是一个既抑制肿瘤细胞生长,又表达刺激免疫系统的蛋白的基因。对该基因的研究具有非常重要的理论和临床意义。国外文献报道大多数是针对hIL-24腺病毒表达载体的研究与开发,对原核表达研究较少。原核表达系统具有良好的可操作性,成本低,可获得大量具有生物学活性的重组蛋白,并且可以进一步开发为应用方便、安全的基因重组药物。本实验利用PCR技术从含有白介素-24基因组中扩增出hIL-24的基因片段(491bp),根据同尾酶的原理成功地构建了hIL-24的原核表达载体pQE30- hIL-24,采用镍凝胶亲和层析的方法获得大量纯化目的蛋白。
[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- 伪 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.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2008
【分类号】：R392;Q78

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