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应用微卫星PCR技术鉴别中国林蛙

Identification of rana temporaria chensinensis by technology of microsatellite PCR
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摘要 目的采用微卫星PCR电泳图谱鉴别中国林蛙长白山亚种及混淆蛙种,进而从基源上控制中药材哈蟆油的质量。方法以东北4种不同产地林蛙基因组DNA为原料,根据5对引物的PCR扩增结果,对4种不同产地的林蛙开展鉴别,并采用荧光标记法对结果进行验证。结果在同种引物作用下,4种林蛙的PCR产物电泳图谱显示,DNA条带大小差异明显;荧光标记验证微卫星分子遗传标记方法可以对东北地区4种蛙属动物进行准确鉴别。结论微卫星方法准确可靠,分子遗传标记技术是一个有效的生物鉴定手段,突破了传统鉴定方法,为道地药材保护和利用提供了有力的参考依据,能够用于鉴别物种种类及新物种的产生。 Objective To identify subspecies and confused frog species of Rana Temporaria Chensinensis in Changbai Mountain by PCR electrophoresis patterns,and then to control the quality of Chinese herbal medicines of toad oil from the base source.Methods Genomic DNAs of Rana Chensinensis from four different origins in the northeast area were used as raw materials,based on PCR amplification results of five primers,four Rana Chensinensis of different origins were identified,and the results were verified by the method of fluorescence labeling.Results Under the effect ofthe same primer,difference of the four kinds of Rana Chensinensis‘s size of DNA bands displayed on PCR agarose electrophoresis pattern was significant,and the method of fluorescence labeling verifying microsatellite molecular genetic markers could accurately identify four kinds of Rana Chensinensis in the northeast area.Conclusion The method of microsatellite was accurate and reliable,and the method of molecular genetic markers was an effective means of bioassays,which broke the traditional identification methods,provided a strong reference for conservation and utilization of authentic ingredients,and could be used for identification of species and new species.
机构地区 长春中医药大学
出处 《吉林中医药》 2014年第10期1016-1018,共3页 Jilin Journal of Chinese Medicine
关键词 林蛙 微卫星 PCR rana chensinensis microsatellite PCR
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