| 人乳头瘤病毒18型L_1基因果蝇表达系统的构建和鉴定 |
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| 引用本文： | 闫爱丽,郑瑾,来保长,耿宜萍,张润歧,王一理.人乳头瘤病毒18型L_1基因果蝇表达系统的构建和鉴定[J].陕西医学杂志,2007,36(8):943-945. |
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| 作者姓名： | 闫爱丽 郑瑾 来保长 耿宜萍 张润歧 王一理 |
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| 作者单位： | 1. 西安医学院
2. 西安交通大学生命科学与技术学院癌症研究所,西安,710061 |
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| 摘 要： | 目的:利用基因重组技术,构建人乳头瘤病毒18型(HPV18)L1基因果蝇表达质粒。方法:以pUC18-HPV18为模板,运用PCR方法扩增不含核定位序列的HPV18L1DNA片段;PCR产物连接至PGEM-TEasy质粒,并测序鉴定;将测序正确的HPV18L1DNA片段亚克隆到果蝇表达载体pMT/BiP/V5-HisA中;利用酶切及PCR方法鉴定重组质粒。结果:经双酶切及PCR鉴定,证实成功构建重组果蝇表达质粒pMT/BiP/V5-HPV18L1。结论:pMT/BiP/V5-HPV18L1重组果蝇表达质粒为下一步转染果蝇S2细胞及制备HPV18VLPs奠定了基础。
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| 关 键 词： | 乳头状瘤病毒 人 基因转移技术 果蝇蛋白质类 疫苗 病毒体 |
| 修稿时间： | 2007-01-18 |
Construction and identification of drosophila expression vector of protein L1 gene of human papillomavirus type 18 |
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| Yan Aili ,Zheng Jin ,Lai Baochang et al.Construction and identification of drosophila expression vector of protein L1 gene of human papillomavirus type 18[J].Shaanxi Medical Journal,2007,36(8):943-945. |
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| Authors: | Yan Aili Zheng Jin Lai Baochang |
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| Abstract: | Objective: To construct recombinant HPV18L1 expression system in Drosophila S2 cells. Methods: The truncated HPV18 L1 gene fragment (no nuclear-localization sequence)was amplified by PCR from pUC18-HPV18 plasmid ;PCR products were inserted into the pGEMT-Easy vector and identified by sequencing, then the truncated HPV18L1 fragment was cloned into Drosophila Schneider expression vector pMT/BiP/V5-HisA, to construct pMT/BiP/V5-HisA/ HPV18L1 plasmid;The construct was identified with endonuclease cleavage and PCR amplification . Results: the truncated HPV18L1 fragment was correctively amplified and successfully cloned in to pMT/BiP/V5-HisA expression plasmid. Conclusion: The construct of pMT/BiP/V5-HPV18L1 may provide a prerequisite for expression of HPV18L1 VLPs in Drosophila S2 cells in the future. |
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| Keywords: | Papillomavirus human Gene transfer techniques Drosophila proteins Accines virosome |
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