Yao Diwen, Guo Dongchun, Zhang Yingkun, Chen Zhihua, Gao Xiaowen, Xing Guiling, Yang Xiuqin, Wang Xibiao, Di Shengwei, Cai Jiancheng, Niu Buyue
College of Animal Science and Technology, Northeast Agricultural University, Harbin, China.
State Key Laboratory of Veterinary Biotechnology, Harbin Veterinary Research Institute, Chinese Academy of Agricultural Sciences (CAAS), Harbin, China.
Front Vet Sci. 2023 Jan 17;9:1090833. doi: 10.3389/fvets.2022.1090833. eCollection 2022.
Identification of causative genes or genetic variants associated with phenotype traits benefits the genetic improvement of animals. CISH plays a role in immunity and growth, however, the upstream transcriptional factors of porcine and the genetic variations in these factors remain unclear. In this study, we firstly identified the minimal core promoter of porcine and confirmed the existence of STATx binding sites. Overexpression and RT-qPCR demonstrated STAT5A increased transcriptional activity ( < 0.01) and mRNA expression ( < 0.01), while GATA1 inhibited transcriptional activity ( < 0.01) and the following mRNA expression ( < 0.05 or < 0.01). Then, the putative functional genetic variations of porcine were screened and a PCR-SSCP was established for genotype g.508A>C and g.566C>T. Population genetic analysis showed the A allele frequency of g.508A>C and C allele frequency of g.566C>T was 0.61 and 0.94 in Min pigs, respectively, while these two alleles were fixed in the Landrace population. Statistical analysis showed that Min piglets with CC genotype at g.566C>T or Hap1: AC had higher 28-day body weight, 35-day body weight, and ADG than TC or Hap3: CT animals ( < 0.05, < 0.05). Further luciferase activity assay demonstrated that the activity of g.508A>C in the C allele was lower than the A allele ( < 0.05). Collectively, the present study demonstrated that STAT5A positively regulated porcine transcription, and SNP g.566C>T in the was associated with the Min piglet growth trait.
鉴定与表型性状相关的致病基因或遗传变异有助于动物的遗传改良。细胞因子诱导的Src同源蛋白（CISH）在免疫和生长中发挥作用，然而，猪CISH的上游转录因子及其遗传变异仍不清楚。在本研究中，我们首先鉴定了猪CISH的最小核心启动子，并证实了信号转导和转录激活因子5（STAT5）结合位点的存在。过表达和逆转录定量聚合酶链反应（RT-qPCR）表明，STAT5A增加了CISH的转录活性（P<0.01）和信使核糖核酸（mRNA）表达（P<0.01），而GATA结合蛋白1（GATA1）抑制了CISH的转录活性（P<0.01）及其后续的mRNA表达（P<0.05或P<0.01）。然后，筛选了猪CISH的推定功能遗传变异，并建立了针对基因型g.508A>C和g.566C>T的聚合酶链反应-单链构象多态性（PCR-SSCP）方法。群体遗传分析表明，在民猪中，g.508A>C的A等位基因频率和g.566C>T的C等位基因频率分别为0.61和0.94，而这两个等位基因在长白猪群体中是固定的。统计分析表明，在g.566C>T位点具有CC基因型或单倍型1（Hap1）：AC的民猪仔猪在28日龄体重、35日龄体重和平均日增重方面高于具有TC基因型或单倍型3（Hap3）：CT的动物（P<0.05，P<0.05）。进一步的荧光素酶活性测定表明，g.508A>C的C等位基因的活性低于A等位基因（P<0.05）。总体而言，本研究表明STAT5A正向调控猪CISH转录，且CISH中的单核苷酸多态性（SNP）g.566C>T与民猪仔猪生长性状相关。
