摘要 PNPLA3蛋白属于patatin样磷脂酶家族,能够参与脂肪积累和脂肪动员,且该蛋白与肝脏炎症、肝纤维化密切相关。本研究旨在克隆PNPLA3基因序列并进行生物信息学分析,以简州大耳羊为实验对象,屠宰后,采集皮下脂肪组织,Trizol法提取组织总RNA,利用RT-PCR法克隆山羊PNPLA3基因序列,对所获得的序列进行生物信息学分析。结果显示:克隆得到山羊PNPLA3基因序列1564 bq(GenBank登陆号:MN643056),其中CDS区1344bp,5'UTR234bp,3'UTR49bp,编码447个氨基酸残基;山羊PNPLA3氨基酸序列与绵羊(Ovis aries)、牛(Bos taurus)、大鼠(Rattus norvegicus)、小鼠(Mus musculus)和人(Homo sapiens)的氨基酸序列相似性分别达到83.89%、81.21%、56.38%、56.15%、58.42%;山羊PNPLA3蛋白与绵羊亲缘关系最近,而与原鸡(Gallus gallus)的亲缘关系较远;山羊PNPLA3蛋白为疏水性酸性蛋白,大部分由α螺旋和无规则卷曲构成,分别占比44.07%、39.6%,有18个丝氨酸、3个苏氨酸、3个酪氨酸磷酸化位点,无信号肽剪切位点,有2个跨膜结构域,具有1个PNPLA家族同源结构域,STRING数据库检索到PNLIP、PNPLA2、LIPC、PNLIPRP3等蛋白可能与PNPLA3蛋白存在相互作用关系。本实验成功克隆出山羊PNPLA3基因,为进一步阐明PNPLA3基因在调控山羊脂质代谢中的作用及分子机制提供基础资料。 PNPLA3 belongs to patatin like phospholipase family,which is involved in fat accumulation and mobilization,and is closely related to liver inflammation and fibrosis.The aims of this study was to clone PNPLA3 gene and perform bioinformatics analysis to further elucidate the role of PNPLA3 gene in regulating of goat lipid metabolism.After slaughter,the subcutaneous adipose tissue was collected and the total RNA was extracted from Jianzhou Big-Eared goat by Trizol method.The goat PNPLA3 gene sequence was cloned by RT-PCR,and the obtained sequences were bioinformatics analyzed.The results showed that goat PNPLA3 gene sequence was 1564 bq(GenBank No.MN643056),including the CDS region 1344 bp,5’UTR 234 bp,3’UTR 49 bp,encoding 447 amino acid residues.The similarity of goat PNPLA3 amino acid sequences with Ovis aries,Bos taurus,Rattus norvegicus,Mus musculus and Homo sapiens was 83.89%,81.21%,56.38%,56.15%and 58.42%,respectively.Goat PNPLA3 protein is most closely related to sheep and least closely related to Gallus gallus.Goat PNPLA3 protein is hydrophobic acidic protein,most of which consisted ofα-helix and irregular curl,accounting for 44.07%and 39.6%respectively,with 18 serine,3 threonine,3 tyrosine phosphorylation sites,no signal-peptide shear sites,2 transmembrane domains,and 1 PNPLA family homologous domain.The STRING database retrieval to PNLIP,PNPLA2,LIPC,PNLIPRP3 and other proteins may interact with PNPLA3 protein.In conclusion,PNPLA3 gene of goat was successfully cloned in this experiment,which provides basic information for further elucidated the role and molecular mechanism of PNPLA3 gene in regulating the lipid metabolism in goat.
出处 《中国畜牧杂志》 CAS 北大核心 2021年第4期73-78,共6页 Chinese Journal of Animal Science
基金 国家重点研发计划(2016YFC0500709) 四川省科技计划项目(2020JDJQ0010) 中央高校基本科研业务费专项资金项目(2020PTJS16)。
