| 球花石斛的位点特异性PCR鉴别研究 |
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| 引用本文： | 应依,徐红,王峥涛.球花石斛的位点特异性PCR鉴别研究[J].药学学报,2007,42(1):98-103. |
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| 作者姓名： | 应依 徐红 王峥涛 |
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| 作者单位： | 1. 上海中医药大学,中药研究所,中药标准化教育部重点实验室,上海,201203
2. 上海中医药大学,中药研究所,中药标准化教育部重点实验室,上海,201203;上海中药标准化研究中心,上海,201203 |
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| 基金项目： | 国家自然科学基金;中国APEC科技产业合作基金;上海市科委资助项目;上海市重点学科建设项目 |
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| 摘 要： | 为建立球花石斛的DNA分子标记鉴别方法，本文根据测定及GenBank上登录的109种共计164个石斛样本的rDNA ITS序列，设计了特异性鉴别引物QH-JB1 和QH-JB2，并对球花石斛进行了位点特异性PCR鉴别研究。结果表明，当复性条件为63.5 ℃，1 min时，只有球花石斛的模板DNA能被扩增出约300 bp的阳性扩增带，而其他种石斛均为阴性。证明用本法鉴别球花石斛简便、省时，也适用于干燥球花石斛药材的鉴别，具有广泛的应用前景。
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| 关 键 词： | 球花石斛 石斛 位点特异性PCR 鉴别 |
| 文章编号： | 0513-4870（2007）01-0098-06 |
| 收稿时间： | 2006-03-20 |
| 修稿时间： | 2006-03-20 |
Allele-specific diagnostic PCR authentication of Dendrobium thyrsiflorum |
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| YING Yi,XU Hong,WANG Zheng-tao.Allele-specific diagnostic PCR authentication of Dendrobium thyrsiflorum[J].Acta Pharmaceutica Sinica,2007,42(1):98-103. |
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| Authors: | YING Yi XU Hong WANG Zheng-tao |
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| Institution: | Key Laboratory of Standardization of Chinese Medicines of the Ministry of Education, Institute of Chinese Materia Medica, Shanghai University of Traditional Chinese Medicine, Shanghai 201203, China. |
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| Abstract: | The aim of this study was to establish a diagnostic PCR method for authentication of D. thyrsiflorum Rchb. f. Based on rDNA ITS sequences of the 164 samples from 109 Dendrobium species sequenced and quoted from GenBank, the allele-specific diagnostic primers QH-JB1 and QH-JB2 for authentication were designed. Using the primers, diagnostic PCR was performed with the total DNA of D. thyrsiflorum Rchb. f. and other D. species as templates to establish the positive PCR condition for D. thyrsiflorum Rchb. f. A DNA fragment about 300 bp was amplified from D. thyrsiflorum Rchb. f. with anneal temperature at 63.5 degrees C and anneal time at 1 min, whereas no other DNA fragment was amplified from the rest D. species. The allele-specific diagnostic PCR was proved to be a simple and quick identification technique, suitable for the authentication of both fresh and dried materials of D. thyrsiflorum Rchb. f. |
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| Keywords: | Dendrobium thyrsiflorum Dendrobium allele-specific diagnostic PCR authentication |
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