摘要
为探究猪前纤维蛋白1(Profilin-1,PFN1)基因多态性及其在猪骨骼肌组织中的mRNA表达规律,以藏猪(Tibet pigs, TP)和大约克猪(Yorkshire pigs, YP)作为试验动物,采用Sanger测序技术对PFN1基因CDS区、5’UTR启动子区进行单核苷酸多态性(Single nucleotide polymorphism, SNP)的筛选与基因分型,预测分析了PFN1基因的SNPs位点突变前后转录因子的功能特性;采用RT-qPCR技术检测PFN1基因在30、90、180日龄三个时期TP和YP的腿肌与背最长肌中的mRNA表达量。结果表明:PFN1基因启动子区域存在三个SNPs,且TP与YP基因频率差异显著;产生了8个新增转录因子结合位点,这些位点调控骨骼肌生长发育、细胞形态建成和保持等过程;PFN1基因表达水平于30日龄时最高,随日龄的增加而下降,90、180日龄时TP的PFN1基因表达量均显著(P<0.05)或极显著(P<0.01)低于YP。综上表明,猪PFN1基因SNPs位点多态性与骨骼肌mRNA的差异表达相关,推测PFN1基因对猪早期骨骼肌生长发育与肌纤维细胞形态保持起重要的调控作用,本研究结果为后续进一步探索其分子作用机制奠定基础。
In order to investigate the polymorphism sites of PFN1(Profilin-1)gene and the expression patterns of its mRNA in pig skeletal muscle tissues,the experiment chose Tibet pigs(TP)and Yorkshire pigs(YP)as experimental animals and performed single nucleotide polymorphism(SNP)screening and genotyping in the CDS and 5'UTR regions of PFN1 gene by Sanger sequencing technology,so as to predict and analyze the functional characteristics of transcription factors before and after SNPs mutation of PFN1 gene.RT-qPCR was adopted to detect the mRNA expression levels of PFN1 gene in the leg muscle and longissimus dorsi muscle of TP and YP at 30-day,90-day,180-day old respectively.The results showed that there existed three SNPs in the promoter regions of PFN1 gene and the gene frequency between TP and YP was significantly different.Moreover,eight new transcription factor binding sites were generated,which were closely related to the regulation of processes including the growth of skeletal muscle and the morphogenesis and maintenance of cell.PFN1 gene achieved the highest expression level at 30 days old and had an overall downward trend with the age,and its expression quantity in the TP at 90 and 180 days old was significantly(P<0.05)or extremely(P<0.01)lower than that of YP.In summary,the polymorphism of SNPs sites of PFN1 gene in pigs are associated with the differential expression of mRNA in skeletal muscle,which suggests that PFN1 gene may play a critical role in the early development of skeletal muscle in pigs as well as in the maintenance of muscle fibers,and these findings lay the foundation for the further exploration on its molecular mechanisms.
作者
李新荻
王宇
卢鸿
罗布白珍
何辰庆
段梦琪
叶幼荣
张健
商鹏
LI Xindi;WANG Yu;LU Hong;LUOBUBAIZHEN;HE Chenqing;DUAN Mengqi;YE Yourong;ZHANG Jian;SHANG Peng(College of Animal Sciences,Tibet Agricultural and Animal Husbandry University,Linzhi 860000,China;The Province and Ministry Co-founded Collaborative Innovation Center of Tibet Characteristic Agricultural and Animal Husbandry Resources Research and Development,Linzhi 860000,China;Animal Husbandry and Veterinary Station of Bayi District Linzhi City,Linzhi 860000,China)
出处
《家畜生态学报》
北大核心
2023年第11期10-15,共6页
Journal of Domestic Animal Ecology
基金
西藏自治区重大科技专项项目(XZ202101ZD0005N)
中国农业大学基本科研业务费专项资金项目(2021TC002)
四川省区域创新合作项目(2020YFQ0029)
中央引导地方项目(YDZX20195400004426)。
