Paek Hyo-Jin, Li Zhou-Yan, Quan Biao-Hu, Yin Xi-Jun
Jilin Provincial Key Laboratory of Transgenic Animal and Embryo Engineering, Yanbian University, Yanji, Jilin, China.
Anim Biotechnol. 2023 Dec;34(7):2231-2239. doi: 10.1080/10495398.2022.2083628. Epub 2022 Jun 13.
Knockout of the 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 mutant pig genotyping. To accomplish this, conserved oligonucleotide primer and restriction site were deduced according to the mutated sequence of the mutant pigs. PCR amplification yielded a 176 bp band for all homozygous mutant (), heterozygous mutant () and wild-type (WT) pigs. However, samples produced two fragments with 176 and 87 bp, and WT samples produced one fragment with 87 bp after being digested by . samples were not digested by and yielded a 176 bp band. Thus, we were able to determine the genotype of all pigs using restriction enzyme-mediated PCR-RFLP method. Overall, the present study reported a simple and fast PCR-RFLP genotyping method for mutant pig breeding. The present study may contribute to the establishment of commercial breeding systems and the production of double muscle pigs.
该基因的敲除与舌头增大有关，并且会导致动物哺乳困难。哺乳困难对动物死亡率有严重影响。因此，需要特别护理以确保它们的生存能力。在此，出生后及时确定所有猪的基因型至关重要。本研究的主要目的是开发用于突变猪基因分型的限制性内切酶介导的PCR-RFLP检测方法。为实现这一目标，根据突变猪的突变序列推导保守的寡核苷酸引物和限制性位点。PCR扩增对所有纯合突变体（）、杂合突变体（）和野生型（WT）猪产生一条176 bp的条带。然而，样品经消化后产生176和87 bp的两个片段，WT样品产生一个87 bp的片段。样品未被消化并产生一条176 bp的条带。因此，我们能够使用限制性内切酶介导的PCR-RFLP方法确定所有猪的基因型。总体而言，本研究报道了一种用于突变猪育种的简单快速的PCR-RFLP基因分型方法。本研究可能有助于建立商业育种系统和生产双肌猪。
