College of Animal Science and Technology, Henan University of Animal Husbandry and Economy, No. 6 Longzi North Road, Zhengdong New District, Zhengzhou, 450046, Henan, China.
Henan Key Laboratory of Unconventional Feed Resources Innovative Utilization, Henan University of Animal Husbandry and Economy, Zhengzhou, Henan, China.
Mol Biotechnol. 2024 Sep;66(9):2262-2272. doi: 10.1007/s12033-023-00857-1. Epub 2023 Aug 25.
Limited research has been conducted on porcine miR-155 promoters, and previous study from our group have identified two haplotypes (TT and CC) in different pig breeds, each associated with five fully linked mutation sites within or near the miR-155 gene (Li et al. Dev Comp Immunol 39(1):110-116, 2013). In this study, the promoter region of porcine miR-155 was screened, and two important transcription factors, Foxp3 and RelA, were identified. The binding ability of Foxp3 protein was found to be affected by the first mutation site (A/C) using EMSA analysis. In vitro experiments revealed that the expression level of miR-155 was significantly higher in the C haplotype compared to the T haplotype. Additionally, northern blotting assays indicated that both the first mutation site (A/C) and the fourth mutation site (G/T) had a significant impact on miR-155 expression levels. These findings provide further insights into the transcriptional regulation of porcine miR-155 and identify crucial mutation sites that influence miR-155 expression. This knowledge can serve as a basis for identifying potential molecular markers associated with disease resistance in swine.
关于猪 miR-155 启动子的研究较少，本课题组先前的研究在不同猪品种中鉴定出两种单倍型（TT 和 CC），它们分别与 miR-155 基因内或附近的五个完全连锁的突变位点相关联（Li 等人，Dev Comp Immunol 39(1):110-116, 2013）。在本研究中，筛选了猪 miR-155 的启动子区域，鉴定出两个重要的转录因子 Foxp3 和 RelA。使用 EMSA 分析发现 Foxp3 蛋白的结合能力受到第一个突变位点（A/C）的影响。体外实验表明，C 单倍型的 miR-155 表达水平明显高于 T 单倍型。此外， northern 印迹分析表明，第一个突变位点（A/C）和第四个突变位点（G/T）都对 miR-155 的表达水平有显著影响。这些发现进一步深入了解了猪 miR-155 的转录调控，并鉴定出影响 miR-155 表达的关键突变位点。这些知识可以作为鉴定与猪疾病抗性相关的潜在分子标记的基础。
