| 结核分枝杆菌mpt64基因稳定表达细胞系的建立 |
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| 引用本文： | 柏银兰,王丽梅,杨芳,薛莹,樊爱琳,康健,张薇,何俊杰,徐志凯. 结核分枝杆菌mpt64基因稳定表达细胞系的建立[J]. 科学技术与工程, 2009, 9(8) |
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| 作者姓名： | 柏银兰 王丽梅 杨芳 薛莹 樊爱琳 康健 张薇 何俊杰 徐志凯 |
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| 作者单位： | 第四军医大学基础部微生物学教研室;基础部教学实验中心;西京医院放疗科;西京医院检验科,西安,710032 |
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| 摘 要： | 为建立稳定表达结核分枝杆菌分泌蛋白MTP64的P815细胞系,将mpt64基因克隆到真核表达载体pcDNA3.1(-),构建mpt64真核表达载体.重组质粒酶切鉴定正确后,以Lipofectamine2000转染P815细胞,通过G418筛选后,得到阳性克隆,分别以RT-PCR方法检测mRNA表达,间接免疫荧光和Western-blot检测目的蛋白的表达.表达结核分枝杆菌分泌蛋白MPT64的真核表达载体转染细胞后,RT-PCR可扩增出目的基因片段,转染细胞间接免疫荧光检测阳性,Western-blot检测在约26 kDa处有特异显色条带,证实转染细胞表达目的基因.成功地建立mpt64稳定表达细胞系,为进一步结核疫苗评价奠定一定基础.
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| 关 键 词： | 结核分枝杆菌 mpt64 细胞 表达 |
Construction of Stable Transfected Cell Lines for Expressing Mycobacterium tuberculosis mpt64 Gene |
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| BAI Yin-lan,WANG Li-mei,YANG Fang,XUE Ying,FAN Ai-lin,KANG Jian,ZHANG Wei,HE Jun-jie,XU Zhi-kai. Construction of Stable Transfected Cell Lines for Expressing Mycobacterium tuberculosis mpt64 Gene[J]. Science Technology and Engineering, 2009, 9(8) |
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| Authors: | BAI Yin-lan WANG Li-mei YANG Fang XUE Ying FAN Ai-lin KANG Jian ZHANG Wei HE Jun-jie XU Zhi-kai |
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| Affiliation: | Department of Microbiology;Basic medicine teaching lab center1;Department of radiotherapy;Xi jing Hospital2;Department of clinical laboratory;Xijing Hospital3;Fourth Medical University of PLA;Xi'an 710032;P.R.China |
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| Abstract: | To construct stable transfected cell lines for expressing Mycobacterium tuberculosis mpt64 gene,the mpt64 gene of Mycobacterium tuberculosis was cloned into the eukaryotic expression vector pcDNA3.1(-).After analyzed by restriction enzyme digestion,the recombinant plasmid was transfected into P815 cells by Lipofectamine2000.The expression was analyzed by RT-PCR,indirect immunofluorescence and Western-blot.The eukaryotic expression vector of mpt64 was confirmed by restriction enzyme digestion.The P815 cell l... |
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| Keywords: | mpt64 |
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