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張麗,劉良科,鄭麗平,劉秤利,劉虹,覃瑞.野葛特異性PCR檢測方法的建立與應用研究[J].中南民族大學學報自然科學版,2019,(2):204-209
野葛特異性PCR檢測方法的建立與應用研究
Establishment and application of a specific PCR assay for pueraria lobata detection
  
DOI:10.12130/znmdzk.20190210
中文關鍵詞: 野葛  特異性PCR  查爾酮合成酶基因  成分鑒定
英文關鍵詞: Pueraria lobata, specific PCR, chalcone synthase gene, component identification
基金項目:湖北省自然科學基金資助項目( 2016CFB616) ; 中央高?;究蒲袠I務費專項資金資助項目( CZD18005) ; 湖北省重點實驗室建設基金( 2018BFC360)
作者單位
張麗1,劉良科2,鄭麗平1,劉秤利1,劉虹1,覃瑞1* 1 中南民族大學 生命科學學院/武陵山區特色資源植物種質保護與利用湖北省重點實驗室,武漢 430074;2 懷化學院 生物與食品工程學院,懷化 418008 
摘要點擊次數: 188
全文下載次數: 177
中文摘要:
      一個物種特有的DNA序列可以開發成能夠識別該物種及其加工產品的標記. 為尋找野葛(Pueraria lobata)特有的DNA序列,建立野葛特異性PCR檢測方法,本研究通過對GenBank中野葛基因信息的檢索分析,篩選出同源序列少特異性強的野葛查耳酮合成酶基因作為研究對象. 對該基因的特定區域設計引物,建立了野葛特異性定性PCR檢測方法. 利用該方法對野葛14個不同居群的DNA進行PCR擴增,均能擴增出226 bp的片段. 而用相同引物分別對20種其他植物的DNA進行擴增均未出現特異性擴增產物. PCR靈敏度實驗結果表明該方法的檢測靈敏度達到0.02 ng基因組DNA,對市場上的葛根產品進行檢測結果表明該方法可以有效鑒定葛根產品中是否含有野葛成分. 該野葛特異性PCR檢測方法特異性強、靈敏度高,可用于野葛成分的檢測與鑒定.
英文摘要:
      A species-specific DNA sequence can be developed to identify the species and processed products derived from this species. In order to find out the unique DNA sequence of kudzu (Pueraria lobata), and establish a specific PCR assay for kudzu detection, the gene information of kudzu in GenBank were searched and analyzed by Blastn. The Kudzu chalcone synthase gene with less specific homologous sequences was selected as the research object. By designing primers for a specific region of this gene, a kudzu-specific qualitative PCR detection method was established. PCR amplification with DNA from 14 different kudzu populations could amplify the same 226 bp fragment. And no amplification was observed with DNA from 20 other plants. The sensitivity of this method was assessed to be 0.02 ng of kudzu genomic DNA.Kudzu-derived products test results showed that the method can identify whether the product contains kudzu component. In conclusion, this PCR method proved to have high specificity and sensitivity. It can be used for the detection and identification of Kudzu-derived products.
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