利用限制性片段长度多态性技术鉴别梅花鹿骨种属来源的研究
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摘要
目的:建立一种基于限制性片段长度多态性技术鉴别梅花鹿骨粉的种属来源的方法。方法:对脱钙后的动物骨粉样品进行核糖核酸提取,通过聚合酶链式反应对梅花鹿特征片段进行扩增,并通过限制性片段长度多态性分析进一步对其种属来源进行确证。结果:通过对特征片段进行聚合酶链式反应扩增,可将梅花鹿及马鹿来源的骨粉样品与牛、猪、狗等动物骨粉样品进行区分。通过对限制性内切酶XbaI进行酶切后的片段长度进行分析,可进一步区分梅花鹿与马鹿来源的骨粉样品。结论:建立了一种基于限制性片段长度多态性技术的梅花鹿骨粉种属来源的鉴别方法。
Objective: To establish a method of restriction fragment length polymorphism( RFLP) to determine the origin of sika deer bones. Methods: The DNA in the bone samples was extracted after decalcification,and then amplified using polymerase chain reaction( PCR). The origin of the samples was further identified using RFLP analysis. Results: The bone samples of sika deer and red deer could be distinguished from those of pig,bovine and dog by PCR. And the samples of sika deer and red deer could be further distinguished by RFLP through the analysis of the length of restriction enzyme XbaI. Conclusion: A RFLP method is established to determine the origin of sika deer bones.
Objective: To establish a method of restriction fragment length polymorphism( RFLP) to determine the origin of sika deer bones. Methods: The DNA in the bone samples was extracted after decalcification,and then amplified using polymerase chain reaction( PCR). The origin of the samples was further identified using RFLP analysis. Results: The bone samples of sika deer and red deer could be distinguished from those of pig,bovine and dog by PCR. And the samples of sika deer and red deer could be further distinguished by RFLP through the analysis of the length of restriction enzyme XbaI. Conclusion: A RFLP method is established to determine the origin of sika deer bones.
引文
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11 Vlach J,Javurkova B,Karamonova L,et al.Novel PCR-RFLP system based on rpo B gene for differentiation of Cronobacter species[J].Food Microbiol,2017,62(4):1-8
2 刘国辉,张玉,杨述华.鹿瓜多肽临床应用的疗效观察及分析[J].中国医院药学杂志,2004,24(8):44-45
3 赵静雪,陈敏,崔光红,等.鹿骨类药材DNA提取方法研究[J].中国中药杂志,2011,36(3):370-374
4 中国药典[S].2015年版.二部.附录X
5 何永盛,林霖,赖心田,等.PCR-DGGE技术应用于肉制品属性鉴别研究[J].食品工业,2016,37(2):4-6
6 唐修君,樊艳凤,贾晓旭,等.基于线粒体16SrRNA基因鉴别畜禽肉中5种动物源性成分[J].食品科技,2015,40(12):86-89
7 朱业培,王玮,吕青骎,等.基于基因芯片技术检测6种动物源性成分[J].南京农业大学学报,2015,38(6):1003-1008
8 王自强,何素婷,邵泓,等.肝素钠中猪、牛及羊源性成分的PCR检测[J].中国药师,2015,18(9):1506-1511
9 何素婷,段徐华,邵泓,等.肝水解肽样品中牛源及猪源性成分的PCR检测[J].药物分析杂志,2012,32(6):1064-1068
10 陈颖,吴亚君,徐宝梁,等.根据线粒体保守序列检测鹿属动物源性成分[J].畜牧与兽医,2004,36(5):1-4
11 Vlach J,Javurkova B,Karamonova L,et al.Novel PCR-RFLP system based on rpo B gene for differentiation of Cronobacter species[J].Food Microbiol,2017,62(4):1-8