| 结核分支杆菌Rv0901基因置换型打靶载体的构建 |
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| 引用本文： | 邱云青,鲍朗,赵计林,赵明才,张会东.结核分支杆菌Rv0901基因置换型打靶载体的构建[J].四川生理科学杂志,2003,25(1):21-24. |
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| 作者姓名： | 邱云青 鲍朗 赵计林 赵明才 张会东 |
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| 作者单位： | 四川大学华西医学中心感染免疫室,成都,610041 |
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| 摘 要： | 目的：对结核杆菌H37Rv的Rv0901基因进行体外扩增，构建基因打靶载体，利用基因打靶技术进行结核分支杆菌基因Rv0901功能的研究。方法：结核分支杆菌标准毒力株H37Rv体外培界，扩增目的基因，连接载体及目的片段，切除目的基因，筛选阳性克隆，酶切鉴定。结果：经酶切鉴定PCR产物及插入片段大小与预期值相符，鉴定证实PCR产物及插入片段为所需目的基因片段，成功切除靶片段。证实标记基因插入片段插入方向正确。结论：成功构建了用于结核分支杆菌基因打靶的置换型载体，为随后将进行的Rv0901基因敲除株的建立，Rv0901基因功能的研究奠定了基础。
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| 关 键 词： | 结核分支杆菌 Rv090l基因 基因敲除 基因打靶 |
Construction of the targeting vector of Rv0901 gene in Mycobacterium Tuberculosis |
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| Qiu Yunqing,Bao Lang,Zhao Jilin,Zhao Mingcai,Zhang Huidong Research.Construction of the targeting vector of Rv0901 gene in Mycobacterium Tuberculosis[J].Sichuan Journal of Physiological Sciences,2003,25(1):21-24. |
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| Authors: | Qiu Yunqing Bao Lang Zhao Jilin Zhao Mingcai Zhang Huidong Research |
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| Institution: | Qiu Yunqing,Bao Lang,Zhao Jilin,Zhao Mingcai,Zhang Huidong Research Department of Infection and Immunity,Medical Center in West China,SiChuan University Chengdu 610041) |
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| Abstract: | Object: In order to investigate the function of Rv0901 gene in Mycobacterium tuberculosis,a fragment of Mycobacterium tuberculosis was amplified and targeting vector was constructed. Methods: Culturing the Mycobacterium tuberculosis in vitro, extracting the genome DNA,amplifying the targeted gene with PCR, constructing the targeting vector and identifying with restricted enzymes. Results: The fragment was amplified successfully.The replacement vector with deleted Rv0901 gene was constructed. Conclusion: Constructing successfully the replacement vector which is used to the gene knockout in Mycobacterium Tuberculosis. |
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| Keywords: | Gene knockout Gene targeting Rv0901 gene Mycobacterium tuberculosis |
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