浙龟甲的PCR鉴别研究

Authentication of Clemmys Mutica (Cantor) by PCR

关键词:  浙龟甲  黄喉水龟  线粒体DNA  细胞色素b  PCR

Key words:  Zheguijia  Clemmys mutica (Cantor)  mtDNA  cytochrome b gene  PCR

摘要:

目的 建立聚合酶链式反应（PCR）技术鉴别浙龟甲[来源为龟黄喉水龟Clemmys mutica（Cantor）的背甲及腹甲]真伪的方法。方法 提取浙龟甲及其混淆品的基因组DNA，从NCBI网站上检索黄喉水龟及其混淆品的线粒体基因，进行分析比对，根据黄喉水龟细胞色素b（cytochrome b gene，Cytb）基因上的特异位点应用Oligo软件设计引物，对浙龟甲及其混淆品样本进行PCR扩增。结果 所设计的引物能特异性扩增黄喉水龟，扩增片段长度为357 bp，能有效区分浙龟甲及其混淆品种。结论 建立的PCR技术鉴别浙龟甲方法，具有特异性高、方法简便快捷、实用性较强的特点，在快速准确鉴别浙龟甲方面具有良好的应用前景。

Abstract:

OBJECTIVE To develop a new polymerase chain reaction(PCR) technology for identifying the authenticity of Zheguijia[the dorsal and abdominal carapaces of Clemmys mutica (Cantor)]. METHODS Extract genomic DNA from Zheguijia and its adulterants was extracted. A series of sequences from mtDNA of Zheguijia and its adulterants were downloaded from the GenBank for analysis and comparison. And the Oligo software was used to design a set of primers, according to the specific sites on Cytb genes of Zheguijia. PCR technology was performed out to amplify the specific fragment of Zheguijia and its adulterants. RESULTS The designed specific primers were only used to amplify the genuine products of Zheguijia and the amplified fragments were 357 bp, which could effectively distinguished Zheguijia and its adulterants. CONCLUSION The PCR assay proposed in this study can be used for the quality control of Zheguijia, which is specificity, sensitivity and applicability.