Identification of Tps2 Used as an Endogenous Reference Gene in Qualitative and Real-time Quantitative PCR Detection of Flammulina velutipes
Identification of Tps2 Used as an Endogenous Reference Gene in Qualitative and Real-time Quantitative PCR Detection of Flammulina velutipes
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金针菇定性和实时定量PCR检测中内参基因Tps2的鉴定
DOI：
10.1007/s12161-021-02043-y
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发表时间：
2021-05
影响因子：
2.9
通讯作者：
Shang Ying
中科院分区：
文献类型：
--
作者：
Zhang Chen;Lei Zhan;Li Yinjiao;Yi Lunzhao;Shang Ying
在检测食品掺假的方法中，内源性参考基因的PCR扩增技术日益重要。本文采用改良的SDS-CTAB法提取金针菇DNA，筛选出适合于鉴定农业重要真菌的内源参考基因tre 6P磷酸酶基因（Tps 2）。通过PCR方法检测金针菇和副产物。Tps 2基因在2个F. velutipes和9个non-F.用定性和定量PCR方法对绒螯蟹进行检测，在非绒螯蟹中均未捕获到扩增子。绒柄类SYBR绿色定量PCR分析结果表明，目的Tps 2基因在不同的F.绒柄品种在Taqman探针定量PCR分析中，检测限低至3 pg/uL的DNA模板。综上所述，Tps 2基因是一种可靠、方便、准确的检测F.因此，在加工食品，如酱料和饼干中，也含有大量的金针菇成分。
Among the methods of detecting food adulteration, the PCR amplification techniques of the endogenous reference gene are increasingly significant. In this article, the improved SDS-CTAB is chosen for the DNA extraction of Flammulina velutipes, the tre6P Phosphatase gene (Tps2) is selected an endogenous reference gene suitable to identify agricultural important fungus F. velutipes and by-products via PCR methods. The Tps2 gene was assayed in 2 varieties of F. velutipes and 9 non-F. velutipes species by both qualitative and quantitative PCR, no amplicon was captured in the non-F. velutipes species. By SYBR Green quantitative PCR, the melting curve showed that the target Tps2 sequence displayed high homogeneity in different F. velutipes varieties. In Taqman probe quantitative PCR analysis, the detection limit was as low as 3 pg/uL of DNA template. From what is said in the foregoing, we can conclude that the Tps2 gene is a reliable, convenient, and accurate endogenous reference gene for detecting F. velutipes component, even in the processed food products, such as sauce and biscuits.
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影响因子：
4.1
作者：
Mac Mahon, Shaun;Begley, Timothy H.;Stromgren, Selen A.
通讯作者：
Stromgren, Selen A.
影响因子：
2
作者：
Chu, Gyo Moon;Yang, Jeong Mo;Song, Young Min
通讯作者：
Song, Young Min
影响因子：
11.2
作者：
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通讯作者：
Jia, Le
影响因子：
1.4
作者：
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通讯作者：
Liang Shi;Tian-Jun Zhang;Chao Xu;Ang Ren;Ailiang Jiang;Han-shou Yu;Mingwen Zhao
影响因子：
3.9
作者：
Palapala, VA;Aimi, T;Morinaga, T
通讯作者：
Morinaga, T