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

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

• Technique and Method • Previous Articles

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)

Abstract

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

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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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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