| 腺病毒反向重复序列ITRs的人工合成与克隆 |
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| 引用本文： | 王昆,葛菁萍,李梅,隋栋,高冬妮,楼庄伟,平文祥.腺病毒反向重复序列ITRs的人工合成与克隆[J].黑龙江大学自然科学学报,2009,26(4). |
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| 作者姓名： | 王昆 葛菁萍 李梅 隋栋 高冬妮 楼庄伟 平文祥 |
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| 作者单位： | 黑龙江大学生命科学学院,微生物黑龙江省高校重点实验室,哈尔滨,150080 |
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| 基金项目： | 教育部留学回国人员启动基金，黑龙江省自然科学基金资助项目，黑龙江省教育厅骨干教师项目资助计划，黑龙江大学杰出青年基金资助项目 |
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| 摘 要： | 为获得高GC含量(72%)的腺伴随病毒AAV反向末端重复序列ITRs(178 bp),以自行设计的4段寡核苷酸引物(约59 bp)为模板,采用融合PCR方法将其人工合成.回收目的片段后,通过TA克隆、PCR和酶切鉴定获得8个阳性转化子.经测序鉴定,得到了5个序列完全正确的克隆.本实验为研究含有ITRs的重组杆状病毒介导外源基因在哺乳动物细胞中表达持续时间的影响奠定了基础.
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| 关 键 词： | 反向末端重复序列 基因合成 |
The synthesis and cloning of Adeno-as-sociated virual inverted terminal repeats |
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| WANG Kun,GE Jing-ping,LI Mei,SUI Dong,GAO Dong-ni,LOU Zhuang-wei,PING Wen-xiang.The synthesis and cloning of Adeno-as-sociated virual inverted terminal repeats[J].Journal of Natural Science of Heilongjiang University,2009,26(4). |
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| Authors: | WANG Kun GE Jing-ping LI Mei SUI Dong GAO Dong-ni LOU Zhuang-wei PING Wen-xiang |
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| Institution: | Key Laboratory of Microbiology;School of Life Science;Heilongjiang University;Harbin 150080;China |
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| Abstract: | The high GC content(78%) Adeno-as-sociated virual inverted terminal repeats(178 bp) was cloned by SOE PCR with 4 oligonucleotides fragments as templates.8 positive clones was identified after the Gel collection,TA cloning,PCR and restrictive enzyme identification of the target DNA.The clones was sequenced and the result shows that 5 of them corresponds to the gene designed which made the way for the study of expression period in mammal cell mediated by recombinant Baculovirus with ITRs. |
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| Keywords: | SOE PCR |
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