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  本文關(guān)鍵詞：B、C基因型乙型肝炎病毒X蛋白對(duì)MDR1基因的反式激活能力不同　出處：《福建醫(yī)科大學(xué)》2008年碩士論文　論文類型：學(xué)位論文

  更多相關(guān)文章： 乙型肝炎病毒 基因型 反式激活 多藥耐藥基因

【摘要】： 目的:研究B、C基因型乙型肝炎病毒(HBV)X蛋白對(duì)多藥耐藥基因1(Multidrug resisitance gene 1,MDR1)反式激活能力的差別。 方法: PCR擴(kuò)增B、C基因型HBV X基因并克隆于pcDNA3.1/HisC載體(重組載體分別為pcDNA3.1/HisC-XB及pcDNA3.1/HisC-XC)。以FuGENE6將重組表達(dá)載體轉(zhuǎn)染HepG2細(xì)胞,采用融合表達(dá)的X-press多肽表位抗體,通過(guò)western blot檢測(cè)目的蛋白在不同時(shí)間段的表達(dá)。PCR擴(kuò)增MDR1基因啟動(dòng)子并克隆于螢火蟲熒光素酶報(bào)告載體pGL3-Basi(c重組載體為pGL-MDR)。pGL-MDR分別與pcDNA3.1/HisC-XB或pcDNA3.1/HisC-XC共轉(zhuǎn)染HepG2細(xì)胞,轉(zhuǎn)染后48小時(shí)裂解細(xì)胞并檢測(cè)胞內(nèi)螢火蟲熒光素酶活性,實(shí)驗(yàn)數(shù)據(jù)以SPSS 11.5軟件分析。 結(jié)果: 1、成功克隆X蛋白表達(dá)載體及MDR1報(bào)告載體。Western blot顯示pcDNA3.1/HisC-XB或pcDNA3.1/HisC-XC在HepG2細(xì)胞中均能表達(dá)HBV X蛋白,以轉(zhuǎn)染后48小時(shí)為最高;2、當(dāng)pGL-MDR報(bào)告質(zhì)粒與X基因重組載體或空載體質(zhì)量比在1:2～1:4范圍內(nèi),螢火蟲熒光素酶在各轉(zhuǎn)染細(xì)胞內(nèi)的活性依次為pcDNA3.1/HisC-XB+pGL-MDR組pcDNA3.1/HisC-XC+pGL-MDR組pcDNA3.1/HisC+pGL-MDR組(對(duì)照組),且存在劑量-效應(yīng)關(guān)系。 結(jié)論: B、C基因型HBV X蛋白對(duì)多藥耐藥基因均有反式激活能力,且B基因型強(qiáng)于C基因型。
[Abstract]:Objective: to study the multidrug resisitance gene 1 (MDR) gene of hepatitis B virus (HBV) resisitance gene X protein (HBVX). MDR1) the difference in transactivation ability. Methods: B was amplified by PCR. C genotype HBV X gene was cloned into pcDNA3.1/HisC vector (. The recombinant vectors were pcDNA3.1/HisC-XB and pcDNA3.1 / HisC-XCn.Recombinant expression vectors were transfected into HepG2 cells by FuGENE6. The fusion expressed X-press peptide epitope antibody was used. The promoter of MDR1 gene was amplified by western blot and cloned into the luciferase report vector pGL3-Basi. C Recombinant vector pGL-MDR).pGL-MDR was cotransfected with pcDNA3.1/HisC-XB or pcDNA3.1/HisC-XC into HepG2 cells respectively. 48 hours after transfection, the cells were lysed and the luciferase activity of firefly was detected. The experimental data were analyzed by SPSS 11.5 software. Results: 1. Successful cloning of X protein expression Vector and MDR1 report Vector. Blot showed that both pcDNA3.1/HisC-XB and pcDNA3.1/HisC-XC could express HBV X protein in HepG2 cells. The highest rate was 48 hours after transfection. 2, when the mass ratio of pGL-MDR report plasmid to X gene recombinant vector or empty vector is in the range of 1: 2 and 1: 4. The luciferase activity of firefly in the transfected cells was in turn pcDNA3.1/HisC-XB pGL-MDR group pcDNA3.1/HisC-XC. PGL-MDR group pcDNA3.1/HisC pGL-MDR group (. Control group). And there is a dose-effect relationship. Conclusion: HBV X protein of genotype B C has transactivation ability to multidrug resistance genes, and genotype B is stronger than genotype C.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2008
【分類號(hào)】：R373

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相關(guān)期刊論文 前3條

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本文編號(hào)：1356504