College of Animal Science and Technology, Nanjing Agricultural University, Nanjing 210095, China.
Department of Animal Sciences, Purdue University, West Lafayette, IN 47907-2054, USA.
Biomolecules. 2020 Nov 12;10(11):1545. doi: 10.3390/biom10111545.
Steroid hormone levels are associated with estrous behavior, which affects timely mating and reproductive efficiency in pigs. 17β-hydroxysteroid dehydrogenase type 14 ( modulates steroid synthesis and metabolism. To identify the functional single nucleotide polymorphisms (SNPs) in the porcine gene, ear tissues from Large White and Mi gilts were collected to extract genomic DNA. Variable lengths of truncated promoter of gene were used to determine the promoter activity by a dual luciferase reporter system. The vector HSD17B14 or HSD17B14 was transfected into porcine granulosa cells (GCs). The core promoter region was identified between -72bp and -218bp. Six of seven SNPs had significant differences of allele frequency between Large White and Mi gilts. The plasmids with the wild genotype AA of rs329427898 maintained a smaller fraction of promoter activity compared with the plasmids with the mutant genotype GG, while the plasmids with wild the genotype TT of rs319864566 had a greater promoter activity than the plasmids with the mutant genotype CC. A missense mutation (Phe73Val) caused changes in the structural dynamics and function of the HSD17B14 protein. The highly expressed HSD17B14 degraded less estradiol into estrone, while the relatively lowly expressed HSD17B14 degraded more estradiol into estrone, suggesting the protein activity of HSD17B14 was greater than that of HSD17B14. Moreover, the HSD17B14 group has a greater apoptosis rate of porcine GCs. The gene could been used as a candidate molecular marker for estrus behavior in pigs.
甾体激素水平与发情行为有关，发情行为会影响猪的适时配种和繁殖效率。17β-羟类固醇脱氢酶 14 型（17β-HSD14）调节类固醇的合成和代谢。为了鉴定猪基因中的功能性单核苷酸多态性（SNP），从大白猪和梅山猪的耳组织中提取基因组 DNA。使用截短的基因启动子来确定启动子活性通过双荧光素酶报告系统。将载体 HSD17B14 或 HSD17B14 转染到猪颗粒细胞（GC）中。鉴定出核心启动子区域在-72bp 到-218bp 之间。7 个 SNP 中有 6 个在大白猪和梅山猪之间存在等位基因频率的显著差异。与突变基因型 GG 的质粒相比，具有野生基因型 AA 的 rs329427898 的质粒保持较小的启动子活性部分，而具有野生基因型 TT 的 rs319864566 的质粒具有比突变基因型 CC 的质粒更大的启动子活性。错义突变（Phe73Val）导致 HSD17B14 蛋白结构动力学和功能发生变化。高度表达的 HSD17B14 将较少的雌二醇降解为雌酮，而相对低表达的 HSD17B14 将较多的雌二醇降解为雌酮，这表明 HSD17B14 蛋白的活性大于 HSD17B14。此外，HSD17B14 组猪 GC 的凋亡率较高。该基因可作为猪发情行为的候选分子标记。
