| r结核分枝杆菌PhoP／PhoR基因重组穿梭表达质粒的构建及鉴定 |
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| 引用本文： | 梁晨,姜晓霞,张锋,王霞,吴芳,章乐,吴江东,张春军,张辉,樊超,庄睿,李文娟,张万江. r结核分枝杆菌PhoP／PhoR基因重组穿梭表达质粒的构建及鉴定[J]. 石河子大学学报(自然科学版), 2013, 0(6): 697-701 |
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| 作者姓名： | 梁晨 姜晓霞 张锋 王霞 吴芳 章乐 吴江东 张春军 张辉 樊超 庄睿 李文娟 张万江 |
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| 作者单位： | [1]石河子大学医学院病理生理学教研室／新疆地方与民族高发病教育部重点实验室,石河子832002 [2]石河子大学医学院第一附属医院功能科,石河子832008 [3]新疆医科大学第五附属医院烧伤整形科,乌鲁木齐830011 [4]新疆阿克苏地区第二人民医院检验科,阿克苏843000 |
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| 基金项目： | 国家自然科学基金项目（81260261、81160192、81260241、81160001）,新疆兵团医药卫生专项项目（2012BA022）,石河子大学高层次人才启动项目（RCZX200922） |
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| 摘 要： | 为构建结核杆菌PhoPR双组分系统PhoP、PhoR基因的重组穿梭质粒pMV361-PhoP、pMV361-PhoR并对其进行鉴定。采用结核分枝杆菌国际标准强毒株H37Rv菌株基因组DNA为模版，利用PCR技术分别扩增PhoP、PhoR基因编码序列，先克隆于T—Vector pMD19（Simple），再亚克隆至大肠杆菌-分枝杆菌穿梭表达载体pMV361，并转化E.coli DH5α感受态细胞，提取所得的阳性克隆子获得重组表达质粒pMV361-PhoP、pMV361-PhoR，通过PCR及双酶切鉴定。结果显示，结核分枝杆菌国际标准强毒株H37Rv菌株基因组DNA中扩增出的PhoP、PhoR基因与GenBank公布的序列一致，经过PCR及双酶切鉴定，表明PhoP、PhoR基因均成功插入了pMV361穿梭载体。由此可知，成功构建了重组穿梭质粒pMV361-PhoP、pMV361-PhoR，为进一步研究PhoPR双组分系统奠定了基础。
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| 关 键 词： | 结核杆菌 PhoP PhoR pMV361 |
Construction and Identification of E. coli-Mycobacterium Recombinant Shuttle Expression Plasmid for PhoP/PhoR of Mycobacterium Tuberculosis |
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| LIANG Chen,JIANG Xiaoxia,ZHANG Feng,WANG Xia,WU Fang,ZHANG Le,WU Jiangdong,ZHANG Chunjun,ZHANG Hui,FAN Chao,ZHUANG Rui,LI Wenjuan,ZHANG Wanjiang. Construction and Identification of E. coli-Mycobacterium Recombinant Shuttle Expression Plasmid for PhoP/PhoR of Mycobacterium Tuberculosis[J]. Journal of Shihezi University(Natural Science), 2013, 0(6): 697-701 |
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| Authors: | LIANG Chen JIANG Xiaoxia ZHANG Feng WANG Xia WU Fang ZHANG Le WU Jiangdong ZHANG Chunjun ZHANG Hui FAN Chao ZHUANG Rui LI Wenjuan ZHANG Wanjiang |
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| Affiliation: | 1 (1 Laboratory of Xinjiang Endemic and Ethnic Diseases/Department of Pathophysiology, School of Medicine, Shihezi University, Shihezi 832002, China ; 2 Functional Department, the first affiliated hospital, School of Medicine, Shihezi University; Shihezi 832002, China; 3 Burn and Plastic Surgery Department, Fifth Affiliated Hospital,Xinjiang Medical University,Urumqi 830011,China; 4 Clinical Laboratory, Second People＇s Hospital, Aksu 843000,China) |
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| Abstract: | To construct and identify the recombinant shuttle plasmid pMV361-PhoP and pMV361-PhoR of Mycobacterium tuberculosis of PhoPR two-component system. PCR was used to amplify PhoP, PhoR from the genomic DNA of the Mycobacterium tuberculosis H37Rv. The gene was inserted initially into T-Vector pMD19 （Simple） ,and then was subcloned into pMV361 and was transformed into E. coli DHSa. We extracted the positive clones to obtain recombinant plasmid and identified them by restriction en-donuclease and PCR.The cloned PhoP and PhoR of Mycobacterium tuberculosis H37Rv were consistent with the sequence in GenBank. The recombinant plasmid pMV361-PhoP and pMV361-PhoR were successfully constructed after restriction endonuclease and PCR. The pMV361-PhoP and pMV361-PhoR were successfully constructed,which laid a foundation for further study. |
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| Keywords: | Mycobacteriurn tuberculosis PhoP PhoR pMV361 |
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