基于核质遗传原理建立多重PCR检测方法鉴定阿胶中马、驴源性成分及皮张种源

doi:10.16288/j.yczz.20-108

遗传 ›› 2020, Vol. 42 ›› Issue (10): 1028-1035.doi: 10.16288/j.yczz.20-108

• 技术与方法 • 上一篇

基于核质遗传原理建立多重PCR检测方法鉴定阿胶中马、驴源性成分及皮张种源

谌阳(), 王文君, 付明, 徐国强, 周翔, 刘榜()

华中农业大学动物科学技术学院,农业动物遗传育种与繁殖教育部重点实验室,武汉 430070

收稿日期:2020-04-20 修回日期:2020-07-10 出版日期:2020-10-20 发布日期:2020-09-16
通讯作者: 刘榜 E-mail:shyang@webmail.hzau.edu.cn;liubang@mail.hzau.edu.cn
作者简介:谌阳,在读硕士研究生,专业方向：动物分子生物学与种源检测。E-mail: shyang@webmail.hzau.edu.cn
基金资助:
国家科技重大专项资助编号(2018ZX08012-001)

A multiplex PCR method based on nuclear and cytoplasmic inheritance to identify the horse and donkey-derived components of Asini Colla Corii and the hide

Yang Shen(), Wenjun Wang, Ming Fu, Guoqiang Xu, Xiang Zhou, Bang Liu()

Key Laboratory of Agricultural Animal Genetics, Breeding and Reproduction of Ministry of Education, College of Animal Science and Technology, Huazhong Agricultural University, Wuhan 430070, China

Received:2020-04-20 Revised:2020-07-10 Online:2020-10-20 Published:2020-09-16
Contact: Liu Bang E-mail:shyang@webmail.hzau.edu.cn;liubang@mail.hzau.edu.cn
Supported by:
Supported by the National Science and Technology Major Project No(2018ZX08012-001)

摘要/Abstract

摘要：

阿胶(Asini Colla Corii)及其原料皮张源性成分的鉴定是对阿胶真实性的一种保障,阿胶生产企业和市场监管部门急需马、驴、骡皮张以及阿胶中源性成分鉴别的有效检测方法。本研究基于马、驴核基因组和线粒体基因组筛选物种特异性DNA序列作为检测靶标,设计马、驴特异性引物,建立了鉴别马、驴、骡皮张以及鉴定阿胶中马和驴源性成分的多重PCR方法。结果显示,本文所建立的方法可用于阿胶源性成分及皮张种源的鉴别,其特异性强,只在目标物种中扩增出目的条带,而非目标物种中没有任何扩增产物,而且灵敏度能达到0.2 ng,可为阿胶生产企业和市场监管部门提供技术支撑。

关键词: 阿胶, 核基因, 线粒体基因, 多重PCR, 马、驴源性成分

Abstract:

To identify the original components of Asini Colla Corii and its raw material hides provides a guarantee for authenticity of Asini Colla Corii. It is urgent for Asini Colla Corii production enterprises and market supervision departments to develop effective identification methods of Asini Colla Corii and hides derived from horses, donkeys, mules and hinnies. This study screened species-specific DNA sequences of nuclear and mitochondrial genomes as detection targets, designed horse and donkey specific primers and established multiple PCR identification methods for identifying the animal hides (including the horse, donkey, mule and hinny) and Asini Colla Corii containing horse-derived and donkey-derived components. Our method can identify the horse, donkey, mule and hinny hides and horse, donkey-derived components of Asini Colla Corii with high species specificity (no crossed amplification was observed ). The limit of detection was 0.2 ng DNA. The method developed in this study provides technical support for Asini Colla Corii production enterprises and market supervision departments.

Key words: Asini Colla Corii, nuclear gene, mitochondrial gene, multiplex PCR, horse and donkey-derived components

谌阳, 王文君, 付明, 徐国强, 周翔, 刘榜. 基于核质遗传原理建立多重PCR检测方法鉴定阿胶中马、驴源性成分及皮张种源[J]. 遗传, 2020, 42(10): 1028-1035.

Yang Shen, Wenjun Wang, Ming Fu, Guoqiang Xu, Xiang Zhou, Bang Liu. A multiplex PCR method based on nuclear and cytoplasmic inheritance to identify the horse and donkey-derived components of Asini Colla Corii and the hide[J]. Hereditas(Beijing), 2020, 42(10): 1028-1035.

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目的基因	引物名称	序列(5?→3?)	目的片段大小(bp)
OBSCN	Horse-NF	CAGGTGAGCCGAGCTGCGCT	166
OBSCN	Horse-NR	TGGCGCACGCAGTTCGTCCC	166
LOC106835644	Donkey-NF	CTGTCCTGATGGAAGAGAACTTGG	144
LOC106835644	Donkey-NR	GGTGGTGCTGCAAATTCCACTTTA	144
ND2	Horse-MF	AAATCCCCTTATCTTCACAACT	179
ND2	Horse-MR	GAAAATATTTGGTGGAGGCTTC	179
ND2	Donkey-MF	CATCCTACTAACTATAGCCGTG	130
ND2	Donkey-MR	CACTAAGATGGCTGCTATTC	130

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基于核质遗传原理建立多重PCR检测方法鉴定阿胶中马、驴源性成分及皮张种源

A multiplex PCR method based on nuclear and cytoplasmic inheritance to identify the horse and donkey-derived components of Asini Colla Corii and the hide

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