Jia Ruike, Xu Lingna, Sun Dongxiao, Han Bo
Department of Animal Genetics and Breeding, College of Animal Science and Technology, Key Laboratory of Animal Genetics, Breeding and Reproduction of Ministry of Agriculture and Rural Affairs, National Engineering Laboratory for Animal Breeding, China Agricultural University, Beijing, China.
National Dairy Innovation Center, Hohhot, China.
Front Genet. 2023 Jan 4;13:1065096. doi: 10.3389/fgene.2022.1065096. eCollection 2022.
homolog, nuclear pore and COPII coat complex component () is the core component of the cytoplasmic COPII complex, which mediates material transport from the endoplasmic reticulum to the Golgi complex. Our preliminary work found that gene was differentially expressed in dairy cows during different stages of lactation, and involved in metabolic pathways of milk synthesis such as citric acid cycle, fatty acid, starch and sucrose metabolisms, so we considered that the might be a candidate gene affecting milk production traits. In this study, we detected the polymorphisms of gene and verified their genetic effects on milk yield and composition traits in a Chinese Holstein cow population. By sequencing the whole coding and partial flanking regions of , we found four single nucleotide polymorphisms (SNPs). Subsequent association analysis showed that these four SNPs were significantly associated with milk yield, fat yield, protein yield or protein percentage in the first and second lactations ( ≤.0351). We also found that two SNPs in formed one haplotype block by Haploview4.2, and the block was significantly associated with milk yield, fat yield, fat percentage, protein yield or protein percentage ( ≤ .0373). In addition, we predicted the effect of SNP on 5'region on transcription factor binding sites (TFBSs), and found that the allele A of 22:g.54362761A>G could bind transcription factors (TFs) GATA5, GATA3, HOXD9, HOXA10, CDX1 and Hoxd13; and further dual-luciferase reporter assay verified that the allele A of this SNP inhibited the fluorescence activity. We speculate that the A allele of 22:g.54362761A>G might inhibit the transcriptional activity of gene by binding the TFs, which may be a cause mutation affecting the formation of milk production traits in dairy cows. In summary, we proved that has a significant genetic effect on milk production traits and the identified significant SNPs could be used as candidate genetic markers for GS SNP chips development; on the other hand, we verified the transcriptional regulation of 22:g.54362761A>G on gene, providing research direction for further function validation tests.
同源物、核孔和COPII包被复合体成分()是细胞质COPII复合体的核心成分,介导从内质网到高尔基体复合体的物质运输。我们的前期工作发现,该基因在奶牛泌乳不同阶段存在差异表达,并参与柠檬酸循环、脂肪酸、淀粉和蔗糖代谢等乳合成代谢途径,因此我们认为该基因可能是影响产奶性状的候选基因。本研究中,我们检测了该基因的多态性,并在中国荷斯坦奶牛群体中验证了它们对产奶量和乳成分性状的遗传效应。通过对该基因的整个编码区和部分侧翼区进行测序,我们发现了4个单核苷酸多态性(SNP)。随后的关联分析表明,这4个SNP在第一和第二泌乳期与产奶量、乳脂产量、乳蛋白产量或乳蛋白率显著相关(P≤0.0351)。我们还发现,该基因中的2个SNP通过Haploview4.2软件形成了一个单倍型块,该单倍型块与产奶量、乳脂产量、乳脂率、乳蛋白产量或乳蛋白率显著相关(P≤0.0373)。此外,我们预测了该基因5′区域SNP对转录因子结合位点(TFBS)的影响,发现22:g.54362761A>G位点的A等位基因可结合转录因子(TF)GATA5、GATA3、HOXD9、HOXA10、CDX1和Hoxd13;进一步的双荧光素酶报告基因试验验证了该SNP的A等位基因抑制荧光活性。我们推测,22:g.54362761A>G位点的A等位基因可能通过结合TF抑制该基因的转录活性,这可能是影响奶牛产奶性状形成的一个致变因素。综上所述,我们证明了该基因对产奶性状具有显著遗传效应,所鉴定的显著SNP可作为GS SNP芯片开发的候选遗传标记;另一方面,我们验证了22:g.54362761A>G对该基因的转录调控作用,为进一步的功能验证试验提供了研究方向。