Application of PCR-RFLP for quick identification of MSTN mutants in MSTN mutant pig breeding

Knockout of the MSTN gene is linked to the enlarged tongue, and it causes suckling difficulty in animals. The suckling difficulty has a severe effect on animal mortality. Thus, special care was required to ensure their survivability. Here, it is critical to promptly ascertain the genotype of all pig...

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Published inAnimal biotechnology Vol. 34; no. 7; pp. 2231 - 2239
Main Authors Paek, Hyo-Jin, Li, Zhou-Yan, Quan, Biao-Hu, Yin, Xi-Jun
Format Journal Article
LanguageEnglish
Published England Taylor & Francis 01.12.2023
Taylor & Francis Ltd
Subjects
Animal husbandry
Breeding
Enzymes
Genotypes
Genotyping
Hogs
Livestock breeding
molecular breeding
MSTN gene
Mutants
myostatin
Oligonucleotides
PCR-RFLP
Polymerase chain reaction
Restriction fragment length polymorphism
sequencing
Suckling behavior
Survivability
Swine
PCR-RFLP
Genotyping
molecular breeding
myostatin
sequencing
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Summary:Knockout of the MSTN gene is linked to the enlarged tongue, and it causes suckling difficulty in animals. The suckling difficulty has a severe effect on animal mortality. Thus, special care was required to ensure their survivability. Here, it is critical to promptly ascertain the genotype of all pigs after birth. The main objective of the present study was to develop the restriction enzyme-mediated PCR-RFLP assay for MSTN mutant pig genotyping. To accomplish this, conserved oligonucleotide primer and restriction site were deduced according to the mutated sequence of the MSTN mutant pigs. PCR amplification yielded a 176 bp band for all homozygous MSTN mutant (MSTN -/- ), heterozygous MSTN mutant (MSTN +/- ) and wild-type (WT) pigs. However, MSTN + /- samples produced two fragments with 176 and 87 bp, and WT samples produced one fragment with 87 bp after being digested by BstNI. MSTN -/- samples were not digested by BstNI and yielded a 176 bp band. Thus, we were able to determine the genotype of all pigs using BstNI restriction enzyme-mediated PCR-RFLP method. Overall, the present study reported a simple and fast PCR-RFLP genotyping method for MSTN mutant pig breeding. The present study may contribute to the establishment of commercial breeding systems and the production of double muscle pigs.
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ISSN:1049-5398
1532-2378
DOI:10.1080/10495398.2022.2083628