| HBX基因重组腺病毒载体的构建 |
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| 引用本文： | 冯涛,何通川,宋文鑫. HBX基因重组腺病毒载体的构建[J]. 重庆医科大学学报, 2004, 29(3): 273-275,278 |
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| 作者姓名： | 冯涛 何通川 宋文鑫 |
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| 作者单位： | 重庆医科大学基础医学院生物化学教研室,重庆,400016;芝加哥大学医学中心分子肿瘤研究室,美国,60637 |
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| 摘 要： | 目的:构建携HBX基因的腺病毒载体,为进一步研究HBX基因的功能,为阐明HBV感染导致肝癌的机制研究建立基础.方法:采用PCR技术从HBV全基因组DNA中扩增HBX基因;将HBX基因亚克隆至质粒pHAHA,构建质粒pHAHA-HBX,酶切pHAHA-HBX质粒,将HAHA-HBX片段克隆到腺病毒载体pAd-Track-CMV中,得到pAdTrack-CMV-HBX,同时与pAdEasy-1共转染293细胞,确定转染效率.检测培养上清病毒中HBX的存在.结果:将PCR扩增HBX片段成功克隆到腺病毒载体中,并经序列分析证实;建立了产病毒细胞株,证实重组病毒中含有HBX基因.结论:成功构建了携HBx基因的腺病毒载体.
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| 关 键 词： | 基因 HBX HBV 腺病毒载体 乙型肝炎 |
| 文章编号： | 0253-3626(2004)03-0273-03 |
Construction of a recombinant adenovirus vector containing HBX gene |
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| FENG Tao,et al. Construction of a recombinant adenovirus vector containing HBX gene[J]. Journal of Chongqing Medical University, 2004, 29(3): 273-275,278 |
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| Authors: | FENG Tao et al |
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| Abstract: | Objective:To establish a recombinant adenovirus vector carrying HBX gene for investigation of the functions of HBX gene on the hepatocellular carcinoma.Methods:HBX gene was amplified from genomic DNA of HBV by polymerase chain reaction(PCR) technique, and subcloned to plasmid pHAHA to construct recombinant plasmid pHAHA-HBX.subcloned HAHA-HBX into a shuttle vector pAdTrack-CMV.After being identified by endonuclease,and subsequently cotransformed into E.coli.293 cells with an adenoviral backbone plasmid,e.g.pAdEasy-1.Results:HBX gene was cloned from HBV genome by PCR successfully,and proved by sequence determination.A vector producing cell line 293/HBX was established.An HBX gene integration was detected by PCR in the recombinant adenovirus vectors.Conclusion:A recombinant adenovirus vector containing HBX gene was successfully constructed. |
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| Keywords: | Gene HBX HBV Adenovirus vector Hepatitis B |
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