| 双自杀基因融合基因重组腺病毒的制备与鉴定 |
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| 引用本文： | 赵军,冯跃,孟荣贵,黄盛东. 双自杀基因融合基因重组腺病毒的制备与鉴定[J]. 第二军医大学学报, 2003, 24(1): 64-66 |
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| 作者姓名： | 赵军 冯跃 孟荣贵 黄盛东 |
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| 作者单位： | 第二军医大学东方肝胆医院胆道二科,上海,200438;长海医院普通外科;长海医院胸心外科 |
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| 基金项目： | 上海市科学技术发展基金 (0 14 1190 41) |
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| 摘 要： | 目的：制备含胞嘧部氨酶（CD）基因,胸苷激酶（TK）基因的融合基因重组腺病毒,为恶性肿瘤的基因治疗研究作准备。方法：构建包含有CD,TK融合基因的重组粘粒,将其与腺病毒DNA末端肽复合物混合后以磷酸钙共沉淀法转染人胚肾293细胞株细胞,获取重组腺病毒,并进行鉴定。结果：酶切结果及PCR结果显示,重组粘粒中CD,TK融合基因插入正确,经鉴定所获重组腺病毒带有融合基因,并且无具有复制能力的腺病毒存在。结论：所获重组病毒为E1,E3缺陷型,含有所需的双自杀基因,可进一步用于基因治疗研究。
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| 关 键 词： | 重组腺病毒 基因疗法 粘粒 胞嘧啶脱氨酶 胸苷激酶 |
| 文章编号： | 0258-879X(2003)01-0064-03 |
| 修稿时间： | 2002-04-22 |
Construction and identification of recombinant adenovirus containing double suicide fusion gene |
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| ZHAO Jun ,FENG Yue ,MENG Rong Gui ,HUANG Sheng Dong. Construction and identification of recombinant adenovirus containing double suicide fusion gene[J]. Former Academic Journal of Second Military Medical University, 2003, 24(1): 64-66 |
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| Authors: | ZHAO Jun FENG Yue MENG Rong Gui HUANG Sheng Dong |
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| Affiliation: | ZHAO Jun 1*,FENG Yue 2,MENG Rong Gui 2,HUANG Sheng Dong 3 |
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| Abstract: | Objective:To construct a recombinant adenovirus containing cytosine deaminase( CD ) gene and thymidine kinase( TK ) fusion gene for the gene therapy research of malignant tumors. Methods: A recombinant cosmid containing CD and TK fusion gene was constructed, and then mixed with DNA TPC and co transfected to the 293 cells by calcium phosphate coprecipitation. Results: The results of restriction and PCR showed the insertion was right and the recombinant adenovirus generated contained the CD and TK fusion gene without replication competent adenovirus. Conclusion: The recombinant adenovirus generated is E1 and E3 deleted and contains double suicide gene needed, which can be further studied for gene therapy. |
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| Keywords: | recombinant adenovirus gene therapy cosmid cytosine deaminase thymidine kinase |
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