| 拮抗链霉菌AFLP分析技术体系的研究(英文) |
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| 引用本文： | 金来武,刘伟成,潘争艳,裘季燕,刘学敏.拮抗链霉菌AFLP分析技术体系的研究(英文)[J].Agricultural Science&Technology,2009(3). |
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| 作者姓名： | 金来武 刘伟成 潘争艳 裘季燕 刘学敏 |
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| 作者单位： | 辽宁省凌源市森林病虫害防治站;北京市农林科学院植物保护环境保护研究所;东北农业大学植保系; |
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| 基金项目： | Supported by the National Natural Science Foundation of China(30671346);;Beijing Natural Science Foundation (6072010);;Beijing Municipal Science and Technology Plan Project (D0706005000091)~~ |
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| 摘 要： | Objective] The aim of this study was to investigate the preparation method and amplification system of antagonistic streptomyces DNA templates based on AFLP assays, and also provide a basis for the application of AFLP technology in the analysis of streptomyces or even actinomyces. Method] The DNAs were extracted by the modified CTAB method and amplified by the Pst Ⅰ/Mse Ⅰ AFLP kit and its reaction system. The amplified products were analyzed by the denatured polyacrylamide gel electrophoresis. Result] The genomic DNAs of ten antagonistic strains of Streptomyces were extracted and tested. The result of 0.8% agarose gel electrophoresis showed that the major DNA bands were clear without degradation and RNA residue, with the fragment sizes ranging from 37.64 to 40.86 Kb. By ultraviolet spectrophotometry, the OD260/OD280 values varying from 1.625 to 1.833 were obtained. Furthermore, the agarose gel electrophoresis of DNA products digested by Pst Ⅰ/Mse Ⅰ presented the dispersed fluorescent long band, which indicated that the enzymatic hydrolysis was fully carried out. The amplified bands of DNA templates by the screened three pairs of primers were clear with rich polymorphism. Conclusion] The preparation method and amplification system of DNA template established in this study can be used in the AFLP analysis of Streptomyces.
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| 关 键 词： | 试剂盒 拮抗链霉菌 森林土壤 异戊醇 菜田土壤 技术体系 双酶切 菌株 琼脂糖凝胶电泳 缓冲液 |
Study on AFLP Technical Protocol for Antagonistic Strains of Streptomyces |
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| JIN Lai-wu,LIU Wei-cheng,PAN Zheng-yan,QIU Ji-yan,LIU Xue-min. Lingyuan Forest Pest , Disease Control Station in Liaoning,Lingyuan.Study on AFLP Technical Protocol for Antagonistic Strains of Streptomyces[J].农业科学与技术(英文版),2009(3). |
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| Authors: | JIN Lai-wu LIU Wei-cheng PAN Zheng-yan QIU Ji-yan LIU Xue-min Lingyuan Forest Pest Disease Control Station in Liaoning Lingyuan |
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| Institution: | JIN Lai-wu1,2,LIU Wei-cheng2,PAN Zheng-yan2,3,QIU Ji-yan2,LIU Xue-min31. Lingyuan Forest Pest , Disease Control Station in Liaoning,Lingyuan 122500,2. Institute of Plant , Environment Protection,Beijing Academy of Agriculture , Forestry Sciences,Beijing 100097,3. Department of Plant Protection,Northeast Agricultural University,Harbin 150030 |
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| Abstract: | Objective] The aim of this study was to investigate the preparation method and amplification system of antagonistic streptomyces DNA templates based on AFLP assays, and also provide a basis for the application of AFLP technology in the analysis of streptomyces or even actinomyces. Method] The DNAs were extracted by the modified CTAB method and amplified by the Pst Ⅰ/Mse Ⅰ AFLP kit and its reaction system. The amplified products were analyzed by the denatured polyacrylamide gel electrophoresis. Result] The genomic DNAs of ten antagonistic strains of Streptomyces were extracted and tested. The result of 0.8% agarose gel electrophoresis showed that the major DNA bands were clear without degradation and RNA residue, with the fragment sizes ranging from 37.64 to 40.86 Kb. By ultraviolet spectrophotometry, the OD260/OD280 values varying from 1.625 to 1.833 were obtained. Furthermore, the agarose gel electrophoresis of DNA products digested by Pst Ⅰ/Mse Ⅰ presented the dispersed fluorescent long band, which indicated that the enzymatic hydrolysis was fully carried out. The amplified bands of DNA templates by the screened three pairs of primers were clear with rich polymorphism. Conclusion] The preparation method and amplification system of DNA template established in this study can be used in the AFLP analysis of Streptomyces. |
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| Keywords: | Antagonistic Streptomyces DNA extraction AFLP |
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