阿胶原料驴源性成分的DNA分子鉴别方法

目的：建立阿胶原料驴源性成分的鉴别方法。方法：通过聚合酶链式反应（PCR），扩增基因组上的线粒体细胞色素b（Cytb）基因，继而用限制性内切酶BamHⅠ对该聚合酶链式反应产物进行酶切，将酶切产物进行琼脂糖凝胶电泳及紫外成像。结果：驴源性成分Cytb的PCR产物能够被BamHⅠ酶切成76 bp和314 bp 2个片段，而马源性成分均不能够被酶切。结论：结合聚合酶链式反应与限制性内切酶片段长度多态性法（PCR-RFLP），可以鉴别阿胶原料驴源性成分。

Objective: To identify donkey-derived components in Ejiao at the DNA level.Methods: The mitochondrial DNA cytochrome b(Cytb)gene was amplified with polymerase chain reaction (PCR). The PCR products were incubated with the restrictive endonuclease BamH Ⅰ, followed by agarose gel electrophoresis and image acquiring under UV light.Results: The PCR products amplified from the Cytb gene of donkey-derived components in Ejiao could be cleaved into two fragments of about 76 bp and 314 bp after incubation with BamH Ⅰ, while the PCR products from horse-derived components could not.Conclusion: A successful identification of donkey-derived components in Ejiao could be achieved by the application of polymerase chain reaction and restriction fragment length polymorphism(PCR-RFLP).

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