Differentiating among horse (Equus caballus), donkey (Equus asinus) and their hybrids with combined analysis of nuclear and mitochondrial gene polymorphism.
Abstract: A novel and brief method of differentiating among horse (Equus caballus) and donkey (Equus asinus) and their hybrids (mule, E. asinus x E. caballus and hinny, E. caballus x E. asinus) with combined analysis of nuclear and mitochondrial gene polymorphism (CANMGP) was reported in the present report. A nuclear gene, protamine P1 gene of donkey was sequenced and compared with the known horse sequence from GenBank while a published equid mitochondrial gene, cytochrome b gene of donkey was compared with that of horse. In each of the two genes, a fixed nucleotide substitution within an exon that could be recognized by Dpn II restriction enzyme was found between the two species. Two pairs of primers were designed for amplifying the fragments within the two genes containing the informative nucleotide positions in 65 horses and 41 donkeys and 38 hybrids and conditions of polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) analysis were optimized. Horse, donkey and mule and hinny had their own specific cleavage patterns after the PCR-RFLP analysis was performed, which made it very easy to identify them from each other. As multiplex PCR can be conducted with the two pairs of primers and only one restriction enzyme is involved in PCR-RFLP analysis, the method described in the present study is a convenient way to identify horse and donkey and their hybrids. The idea involved in the method of CANMGP can be also used to differentiate other animal species or breeds and their hybrids.
Publication Date: 2005-08-03 PubMed ID: 16060497DOI: 10.1111/j.1439-0388.2005.00535.xGoogle Scholar: Lookup
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- Journal Article
Summary
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The research presents a new and efficient method to differentiate between horses, donkeys, and their hybrids using combined analysis of nuclear and mitochondrial gene polymorphism (CANMGP).
Objective and Rationale
- The objective of this study was to develop a simple technique to distinguish horses (Equus caballus) from donkeys (Equus asinus) and their hybrids (mules and hinnies).
- The rationale was based on the idea that conducting a combined analysis of nuclear and mitochondrial gene polymorphism (CANMGP) could differentiate between the species conveniently, providing a unique way of identifying breeds or species based on their gene polymorphisms.
Research Methodology
- The researchers sequenced a nuclear gene, protamine P1, from the donkey and compared it to the known horse sequence from GenBank.
- They also compared a published equid mitochondrial gene, the cytochrome b gene, from the donkey to the horse’s equivalent gene.
- A fixed nucleotide substitution within an exon that could be recognized by the Dpn II restriction enzyme was found between the two species in both genes.
- Primers were prepared to amplify fragments within the two genes containing the informtive nucleotide positions in 65 horses, 41 donkeys, and 38 hybrids.
Outcomes
- The conditions of polymerase chain reaction and restriction fragment length polymorphism (PCR-RFLP) analysis were optimized, generating specific cleavage patterns for horses, donkeys, and their hybrids.
- This identification was facilitated by the multiplex PCR with two pairs of primers and a single restriction enzyme involved in the PCR-RFLP analysis.
Significance
- The study provides a straightforward way to identify horse and donkey species and their hybrids.
- The novel technique also hints at its potential application to differentiate other animal species or breeds and their hybrids by studying gene polymorphisms.
Cite This Article
APA
Zhao CJ, Han GC, Qin YH, Wu Ch.
(2005).
Differentiating among horse (Equus caballus), donkey (Equus asinus) and their hybrids with combined analysis of nuclear and mitochondrial gene polymorphism.
J Anim Breed Genet, 122(4), 285-288.
https://doi.org/10.1111/j.1439-0388.2005.00535.x Publication
Researcher Affiliations
- Equine Center, China Agricultural University, Beijing, China.
MeSH Terms
- Animals
- Cytochromes b / genetics
- DNA / blood
- DNA / chemistry
- DNA / isolation & purification
- Deoxyribonucleases, Type II Site-Specific / metabolism
- Equidae / genetics
- Female
- Genes, Mitochondrial / genetics
- Hair / chemistry
- Horses / genetics
- Hybridization, Genetic / genetics
- Male
- Polymerase Chain Reaction / veterinary
- Polymorphism, Genetic
- Polymorphism, Restriction Fragment Length
- Protamines / genetics
- Restriction Mapping / veterinary
Citations
This article has been cited 6 times.- Zhang J, Wu M, Ma Z, Zhang Y, Cao H. Species-specific identification of donkey-hide gelatin and its adulterants using marker peptides. PLoS One 2022;17(8):e0273021.
- Sheu SC, Huang JY, Lien YY, Lee MS. Specific, sensitive and rapid authentication of donkey-hide gelatine (Colla corii asini) in processed food using an isothermal nucleic acid amplification assay. J Food Sci Technol 2020 Aug;57(8):2877-2883.
- Zuo HL, Zhao J, Wang YT, Xia ZN, Hu YJ, Yang FQ. Identification of the Adulterated Asini Corii Colla with Cytochrome c Oxidase Subunit I Gene-based Polymerase Chain Reaction. Pharmacognosy Res 2017 Oct-Dec;9(4):313-318.
- Han H, Wang A, Liu L, Zhao G, Su J, Wang B, Li Y, Zhang J, Wu B, Sun W, Hu S, Li S, Zhao L, Li X. Testicular Characteristics and the Block to Spermatogenesis in Mature Hinny. Asian-Australas J Anim Sci 2016 Jun;29(6):793-800.
- Millard JT, Chuang E, Lucas JS, Nagy EE, Davis GT. Case-Study Investigation of Equine Maternity via PCR-RFLP: A Biochemistry Laboratory Experiment. J Chem Educ 2013 Nov 12;90(11).
- Parsad R, Bagiyal M, Ahlawat S, Arora R, Gera R, Chhabra P, Sharma U. Unraveling the genetic and physiological potential of donkeys: insights from genomics, proteomics, and metabolomics approaches. Mamm Genome 2025 Mar;36(1):10-24.
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