| 基于重组溶葡球菌酶和ATP生物发光技术特异定量检测金黄色葡萄球菌 |
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| 引用本文： | 李玉元,米志强,安小平,周育森,童贻刚.基于重组溶葡球菌酶和ATP生物发光技术特异定量检测金黄色葡萄球菌[J].生物工程学报,2014,30(8):1283-1290. |
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| 作者姓名： | 李玉元 米志强 安小平 周育森 童贻刚 |
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| 作者单位： | 1 广西医科大学，广西 南宁 530021；2 军事医学科学院微生物流行病研究所 病原微生物生物安全国家重点实验室，北京 100071;2 军事医学科学院微生物流行病研究所 病原微生物生物安全国家重点实验室，北京 100071;2 军事医学科学院微生物流行病研究所 病原微生物生物安全国家重点实验室，北京 100071;1 广西医科大学，广西 南宁 530021；2 军事医学科学院微生物流行病研究所 病原微生物生物安全国家重点实验室，北京 100071;2 军事医学科学院微生物流行病研究所 病原微生物生物安全国家重点实验室，北京 100071 |
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| 基金项目： | 国家自然科学基金 (No. 81072350)，“重大新药创制”科技重大专项“十二五”实施计划 (No. 2011ZX09401-023)，“艾滋病和病毒性肝炎等重大传染病防治”科技重大专项“十二五”实施计划 (No. 2011ZX10004-001)，病原微生物生物安全国家重点实验室开放课题 (No. SKLPBS1113) 资助。 |
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| 摘 要： | 基于重组溶葡球菌酶和ATP生物发光法建立特异定量检测金黄色葡萄球菌的方法。优化设计合成溶葡球菌酶序列,构建重组表达载体pQE30-Lys,转化至大肠杆菌M15并诱导表达,镍柱纯化得到目的蛋白。利用重组溶葡球菌酶和ATP生物发光法特异定量检测金黄色葡萄球菌并与平板计数对比。成功表达了重组溶葡球菌酶,并建立了特异定量检测金黄色葡萄球菌的方法,与平板计数具有显著线性关系。本研究建立的将重组溶葡球菌酶和ATP生物发光法相结合的检测方法操作快捷简单,具有良好的应用前景。
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| 关 键 词： | 金黄色葡萄球菌 重组溶葡球菌酶 ATP生物发光 特异和定量检测 |
| 收稿时间： | 2013/11/12 0:00:00 |
Quantitative specific detection of Staphylococcus aureus based on recombinant lysostaphin and ATP bioluminescence |
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| Yuyuan Li,Zhiqiang Mi,Xiaoping An,Yusen Zhou and Yigang Tong.Quantitative specific detection of Staphylococcus aureus based on recombinant lysostaphin and ATP bioluminescence[J].Chinese Journal of Biotechnology,2014,30(8):1283-1290. |
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| Authors: | Yuyuan Li Zhiqiang Mi Xiaoping An Yusen Zhou and Yigang Tong |
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| Institution: | 1 Guangxi Medical University, Nanning 530021, Guangxi, China; 2 State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China;2 State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China;2 State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China;1 Guangxi Medical University, Nanning 530021, Guangxi, China; 2 State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China;2 State Key Laboratory of Pathogen and Biosecurity, Institute of Microbiology and Epidemiology, Academy of Military Medical Sciences, Beijing 100071, China |
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| Abstract: | Quantification of Staphylococcus aureus is developed based on recombinant lysostaphin and ATP bioluminescence. To produce recombinant lysostaphin, lysostaphin gene was chemically synthesized and inserted it into prokaryotic expression vector pQE30, and the resulting expression plasmid pQE30-Lys was transformed into E. coli M15 for expressing lysostaphin with IPTG induction. The recombinant protein was purified by Ni2+-NTA affinity chromatography. Staphylococcus aureus was detected by the recombinant lysostaphin with ATP bioluminescence, and plate count method. The results of the two methods were compared. The recombinant lysostaphin was successfully expressed, and quantification of S. aureus has been established, which showed a significant linear correlation with the colony counting. The detection method developed has good perspective to quantify S. aureus |
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| Keywords: | Staphylococcus aureus recombinant lysostaphin ATP bioluminescence specific and quantitative detection |
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