| 川芎内生细菌DGGE分析条件的筛选与优化 |
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| 引用本文： | 何冬梅,严铸云,王海,陈彩霞. 川芎内生细菌DGGE分析条件的筛选与优化[J]. 中国实验方剂学杂志, 2015, 21(18): 26-30 |
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| 作者姓名： | 何冬梅 严铸云 王海 陈彩霞 |
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| 作者单位： | 成都中医药大学 中药资源系统研究与开发利用省部共建国家重点实验室培育基地, 中药材标准化省部共建教育部重点实验室, 成都 611137,成都中医药大学 中药资源系统研究与开发利用省部共建国家重点实验室培育基地, 中药材标准化省部共建教育部重点实验室, 成都 611137,成都中医药大学 中药资源系统研究与开发利用省部共建国家重点实验室培育基地, 中药材标准化省部共建教育部重点实验室, 成都 611137,成都中医药大学 中药资源系统研究与开发利用省部共建国家重点实验室培育基地, 中药材标准化省部共建教育部重点实验室, 成都 611137 |
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| 基金项目： | 国家自然科学基金项目(81001610);四川省教育厅基金项目(14ZB0090,12ZB212) |
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| 摘 要： | 目的:针对川芎内生细菌16S rDNA序列进行变性梯度凝胶电泳(DGGE)适宜序列的筛选与电泳条件优化。方法:川芎内生细菌16S rDNA的V3区及V6~V8区分别进行聚合酶链反应(PCR)及巢式PCR以获得各菌种的目标DNA序列。采用DGGE垂直电泳法优化凝胶变性梯度,时间间隔法优化电泳时间,并根据电泳效果对电压和电泳时间组合进行修正。结果:川芎内生细菌16S rDNA V6~V8区序列更能有效排除宿主植物DNA的干扰,获得更加丰富的内生细菌DNA序列。针对该区的DGGE最佳分析条件为变性剂梯度55%~70%(Gel 6%),电泳温度60℃,电压50 V,电泳时间16 h。结论:利用该优化条件能有效分离川芎各内生细菌16S rDNA V6~V8区序列,为川芎内生细菌的群落结构PCR-DGGE分析提供可靠的技术基础,为其他药用植物内生细菌的菌群多样性分析提供参考。
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| 关 键 词： | 川芎 内生细菌 变性梯度凝胶电泳 聚合酶链反应 引物 |
| 收稿时间： | 2015-03-02 |
Screening and Amelioration of DGGE Analysis Conditions for Endophytic Bacteria of Chuanxiong Rhizoma |
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| HE Dong-mei,YAN Zhu-yun,WANG Hai and CHEN Cai-xia. Screening and Amelioration of DGGE Analysis Conditions for Endophytic Bacteria of Chuanxiong Rhizoma[J]. China Journal of Experimental Traditional Medical Formulae, 2015, 21(18): 26-30 |
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| Authors: | HE Dong-mei YAN Zhu-yun WANG Hai CHEN Cai-xia |
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| Affiliation: | State Key Laboratory Breeding Base of Systematic Research, Development and Utilization of Chinese Medicine Resources, Co-founded by Sichuan Province and Ministry of Science and Technology, Key Laboratory of Standardization of Chinese Herbal Medicine, Ministry of Education, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China,State Key Laboratory Breeding Base of Systematic Research, Development and Utilization of Chinese Medicine Resources, Co-founded by Sichuan Province and Ministry of Science and Technology, Key Laboratory of Standardization of Chinese Herbal Medicine, Ministry of Education, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China,State Key Laboratory Breeding Base of Systematic Research, Development and Utilization of Chinese Medicine Resources, Co-founded by Sichuan Province and Ministry of Science and Technology, Key Laboratory of Standardization of Chinese Herbal Medicine, Ministry of Education, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China and State Key Laboratory Breeding Base of Systematic Research, Development and Utilization of Chinese Medicine Resources, Co-founded by Sichuan Province and Ministry of Science and Technology, Key Laboratory of Standardization of Chinese Herbal Medicine, Ministry of Education, Chengdu University of Traditional Chinese Medicine, Chengdu 611137, China |
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| Abstract: | |
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| Keywords: | Chuanxiong Rhizoma endophytic bacterrium denaturing gradient gel electrophoresis polymerase chain reaction primer |
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