乙型肝炎病毒基因（S、X）载体构建表达及功能研究
发布时间：2021-09-09 19:58
目的分别构建乙型肝炎病毒（Hepatitis B Virus，HBV）S基因（包括Pre-S1，Pre-S2和S）和X基因的带FLAG标签的真核表达载体，并进行有效表达，进而通过高通量蛋白质技术获取与X蛋白相互作用的蛋白质复合物，以期为进一步研究乙型肝炎病毒外膜蛋白及X蛋白的功能打下基础。方法以质粒pcDNA3.1-HBV为模板，通过PCR技术扩增HBV基因组Large surface antigen（LS）、Middle surface antigen（MS）、Smallsurface antigen（SS）和X基因片段；将PCR克隆获得的目的基因片段分别插入真核表达载体pCMV-tag2B，构建重组质粒pCMV-tag2B-LS、 pCMV-tag2B-MS、 pCMV-tag2B-SS和pCMV-tag2B-X，并送测序鉴定。将鉴定正确后的目的基因表达载体分别转染293FT细胞，Western blot检测其表达。继而应用FLAG亲和层析柱分离HBX复合体，iTRAQ标记后用串联质谱对其进行鉴定。选择三次实验中只在HBX复合体中鉴定的蛋白质作为进一步功能分析的侯选蛋白。通过免疫共...
【文章来源】：重庆医科大学重庆市
【文章页数】：50 页
【学位级别】：硕士
【部分图文】：
凝胶电泳分离LS、MS、SS、X的PCR产物
图 1.1 凝胶电泳分离 LS、MS、SS、X 的PCR 产物Fig 1.1 PCR product of LS、MS、SS and Xseperated by Agar Gel electrophoresis图 1.2 pCMV-tag2b-LS, pCMV-tag2b-MS,pCMV-tag2b-SS 双酶切后Fig 1.2 pCMV-tag2b-LS, pCMV-tag2b-MS,pCMV-tag2b-SS after digested by restrictionenzyme
图 1.1 凝胶电泳分离 LS、MS、SS、X 的PCR 产物Fig 1.1 PCR product of LS、MS、SS and Xseperated by Agar Gel electrophoresis图 1.2 pCMV-tag2b-LS, pCMV-tag2b-MS,pCMV-tag2b-SS 双酶切后Fig 1.2 pCMV-tag2b-LS, pCMV-tag2b-MS,pCMV-tag2b-SS after digested by restrictionenzyme
【参考文献】：
期刊论文
[1]Adenovirus-expressed preS2 antibody inhibits hepatitis B virus infection and hepatic carcinogenesis[J]. Qian Zhang,Department of Infectious Diseases,Changhai Hospital,Second Military Medical University,Shanghai 200433,China Zhi-Qing Li,Hu Liu,Jia-He Yang,Department of Hepatic Surgery,Eastern Hepatobiliary Surgery Hospital,Second Military Medical University,Shanghai 200438,China. World Journal of Gastroenterology. 2012(04)
[2]Hepatitis B virus X protein promotes proliferation and upregulates TGF-β1 and CTGF in human hepatic stellate cell line,LX-2[J]. Guang-Hui Guo,De-Ming Tan,Ping-An Zhu and Fei Liu Department of Infectious Diseases,Xiangya Hospital,Central South University,Changsha 410008,China. Hepatobiliary & Pancreatic Diseases International. 2009(01)
本文编号：3392694
【文章来源】：重庆医科大学重庆市
【文章页数】：50 页
【学位级别】：硕士
【部分图文】：
凝胶电泳分离LS、MS、SS、X的PCR产物
图 1.1 凝胶电泳分离 LS、MS、SS、X 的PCR 产物Fig 1.1 PCR product of LS、MS、SS and Xseperated by Agar Gel electrophoresis图 1.2 pCMV-tag2b-LS, pCMV-tag2b-MS,pCMV-tag2b-SS 双酶切后Fig 1.2 pCMV-tag2b-LS, pCMV-tag2b-MS,pCMV-tag2b-SS after digested by restrictionenzyme
图 1.1 凝胶电泳分离 LS、MS、SS、X 的PCR 产物Fig 1.1 PCR product of LS、MS、SS and Xseperated by Agar Gel electrophoresis图 1.2 pCMV-tag2b-LS, pCMV-tag2b-MS,pCMV-tag2b-SS 双酶切后Fig 1.2 pCMV-tag2b-LS, pCMV-tag2b-MS,pCMV-tag2b-SS after digested by restrictionenzyme
【参考文献】：
期刊论文
[1]Adenovirus-expressed preS2 antibody inhibits hepatitis B virus infection and hepatic carcinogenesis[J]. Qian Zhang,Department of Infectious Diseases,Changhai Hospital,Second Military Medical University,Shanghai 200433,China Zhi-Qing Li,Hu Liu,Jia-He Yang,Department of Hepatic Surgery,Eastern Hepatobiliary Surgery Hospital,Second Military Medical University,Shanghai 200438,China. World Journal of Gastroenterology. 2012(04)
[2]Hepatitis B virus X protein promotes proliferation and upregulates TGF-β1 and CTGF in human hepatic stellate cell line,LX-2[J]. Guang-Hui Guo,De-Ming Tan,Ping-An Zhu and Fei Liu Department of Infectious Diseases,Xiangya Hospital,Central South University,Changsha 410008,China. Hepatobiliary & Pancreatic Diseases International. 2009(01)
本文编号：3392694
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