【摘要】：慢性粒細(xì)胞白血病(Chronic myeloid leukemia,CML)具有特征性的bcr-abl融合基因。本實(shí)驗(yàn)室成功構(gòu)建表達(dá)bcr-abl基因融合區(qū)內(nèi)大約472bp的融合基因片段的重組表達(dá)載體pcDNAbcr-abl,用其免疫小鼠,能在小鼠體內(nèi)成功誘生針對(duì)bcr-abl融合蛋白的特異性抗體。白細(xì)胞介素-7(Interleukin-7,IL-7)能增強(qiáng)特異性CTL細(xì)胞活性。本課題試圖分三部分研究小鼠IL-7基因?qū)cr-abl融合基因疫苗誘導(dǎo)機(jī)體免疫應(yīng)答的影響。 一、bcr-abl融合基因片段和mlL-7基因雙表達(dá)載體的構(gòu)建與鑒定 為了給小鼠IL-7增強(qiáng)bcr-abl融合基因片段基因疫苗誘導(dǎo)特異性免疫應(yīng)答的研究創(chuàng)造條件,我們將小鼠IL-7基因和bcr-abl融合基因片段分別插入具有兩個(gè)多克隆位點(diǎn)的真核表達(dá)載體pVITR02-mcs,成功構(gòu)建pVbcr-abl/mIL7雙表達(dá)質(zhì)粒。在pVbcr-abl/mIL7質(zhì)粒中,IL-7基因、bcr-abl融合基因片段分別受不同的啟動(dòng)子調(diào)控。我們用pVbcr-abl/mIL7雙表達(dá)載體轉(zhuǎn)染CHO細(xì)胞,分別從蛋白表達(dá)水平和mRNA水平證實(shí),bcr-abl融合基因片段和小鼠IL-7在真核細(xì)胞CHO中得到高效表達(dá)。bcr-abl基因片段和mIL-7基因雙表達(dá)真核細(xì)胞載體的構(gòu)建為慢性粒細(xì)胞白血病bcr-abl融合基因疫苗的研究提供了新的實(shí)驗(yàn)研究工具。 二、兩種bcr-abl基因疫苗對(duì)SP2/0/bcr-abl移植瘤小鼠的影響 在構(gòu)建bcr-abl基因片段和mIL-7基因雙表達(dá)載體、表達(dá)bcr-abl融合基因片段的SP2/0/bcr-abl細(xì)胞的基礎(chǔ)上,我們首先用pVbcr-abl、pVbcr-abl/mIL7兩種質(zhì)粒分別免疫BALB/c小鼠,然后用SP2/0/bcr-abl細(xì)胞攻擊,觀察疫苗對(duì)SP2/0/bcr-abl
[Abstract]:Chronic myeloid leukemia (Chronic myeloid leukemia,CML) has a characteristic bcr-abl fusion gene. In our laboratory, the recombinant expression vector pcDNAbcr-abl, expressing the fusion gene fragment of approximately 472bp in the fusion region of bcr-abl gene was successfully constructed to immunize mice, and the specific antibody against bcr-abl fusion protein was successfully induced in mice. Interleukin-7 (Interleukin-7,IL-7) can enhance the activity of specific CTL cells. This paper attempts to study the effect of mouse IL-7 gene on immune response induced by bcr-abl fusion gene vaccine in three parts. Construction and identification of a bcr-abl fusion gene fragment and a double expression vector of mlL-7 gene in order to induce specific immunity to mouse IL-7 enhanced bcr-abl fusion gene fragment gene vaccine The study of epidemic response creates conditions, The mouse IL-7 gene and the bcr-abl fusion gene fragment were inserted into the eukaryotic expression vector pVITR02-mcs, with two polyclonal sites respectively to construct the pVbcr-abl/mIL7 double expression plasmid. In pVbcr-abl/mIL7 plasmids, the fragment of bcr-abl fusion gene of IL-7 gene was regulated by different promoters. We transfected CHO cells with pVbcr-abl/mIL7 double expression vector. The expression of bcr-abl fusion gene and mouse IL-7 in eukaryotic cell CHO was confirmed by protein expression level and mRNA level, respectively. The eukaryotic expression vector of bcr-abl gene and mIL-7 gene was constructed as bcr-abl in chronic myeloid leukemia. The research of fusion gene vaccine provides a new experimental research tool. Secondly, the effect of two bcr-abl gene vaccines on SP2/0/bcr-abl transplanted tumor mice was based on the construction of bcr-abl gene fragment and mIL-7 gene expression vector, and the expression of bcr-abl fusion gene fragment in SP2/0/bcr-abl cells. We first immunized BALB/c mice with two plasmids of pVbcr-abl,pVbcr-abl/mIL7, then attacked with SP2/0/bcr-abl cells to observe the effect of vaccine on SP2/0/bcr-abl.
【學(xué)位授予單位】：揚(yáng)州大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2005
【分類號(hào)】：R392.1

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