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篇目详细内容 |
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【篇名】 |
Restriction Enzyme Pattern Analysis of Mycobacteria DNA by Capillary Electrophoresis with Laser-Induced Fluorescence Detection |
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【刊名】 |
Frontiers of Chemistry in China |
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【刊名缩写】 |
Front. Chem. China |
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【ISSN】 |
1673-3495 |
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【EISSN】 |
1673-3614 |
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【DOI】 |
10.1007/s11458-005-0002-9 |
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【出版社】 |
Higher Education Press and Springer-Verlag |
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【出版年】 |
2006 |
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【卷期】 |
1
卷1期 |
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【页码】 |
27-33
页,共
7
页 |
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【作者】 |
Li Yuanqian;
Wang Guoqing;
Mi Jianping;
Zhou Ying;
Zeng Hongyan;
Zhang Chaowu;
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【关键词】 |
capillary electrophoresis; laser-induced fluorescence detection; restriction enzyme pattern; Mycobacterium |
【摘要】 |
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A new method for rapidly detecting restriction enzyme patterns of Mycobacterium DNA using capillary electrophoresis with laser-induced fluorescence detection (CE-LIFD) was developed. Polymerase chain reaction was used to amplify a 439-bp fragment of a 65,000-kDa (Mr) heat shock protein gene (hsp65) of Mycobacterium. After digesting amplification products by BstEII and HaeIII, patterns of enzyme cleavage products were detected by both CE-LIFD and agarose gel electrophoresis (AGE), respectively. Experimental parameters of CE were optimized. Restriction enzyme patterns of Mycobacterium DNA were detected in optimum electrophoresis conditions: a coated capillary column with a length of 50 cm and an internal diameter of 100 μm, an electrophoresis buffer of 45 mmol/l Tris–boric acid-ethylenediaminetetraacetic acid, and a running voltage of 11 kV. The restriction enzyme patterns for eight species of mycobacteria were studied. Relative standard deviations of the relative migration times of DNA segments were <3.6%. Compared with AGE, CE is more outstanding in resolution and detection time, and it can be applied as a more effective means to DNA restriction enzyme pattern analysis. |
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