大通牦牛PDGFD基因克隆、生物信息学及组织表达谱分析  

Cloning,Bioinformatics and Tissue Expression Profile Analysis of PDGFD Gene in Yak(Bos mutus)

作　　者：李歆怡 王福彬 黄纯 马晓明 喇永福 褚敏[2] 郭宪[2] 阎萍[2] 潘和平[1] 梁春年[2] Li Xin-Yi;Wang Fu-Bin;Huang Chun;Ma Xiao-Ming;La Yong-Fu;Chu Min;Guo Xian;Yan Ping;Pan He-Ping;Liang Chun-Nian(College of Life Science and Engineering,Northwest Minzu University,Lanzhou 730030,China;Lanzhou Institute of Husbandry and Pharmaceutical Sciences Chinese Academy of Agricultural Sciences,Lanzhou 730050,China)

机构地区：[1]西北民族大学生命科学与工程学院,兰州730030 [2]中国农业科学院兰州畜牧与兽药研究所,兰州730050

出　　处：《农业生物技术学报》2023年第3期518-529,共12页Journal of Agricultural Biotechnology

基　　金：国家重点研发计划(2021YFD1600200);甘肃省科技计划(20JR5RA580);现代农业(肉牛牦牛)产业技术体系建设专项资金(CARS-37);中国农业科学院科技创新工程(25-LZIHPS-01)。

摘　　要：牦牛(Bos mutus)是高原地区十分重要的种质资源。为研究牦牛血小板源性生长因子D(plateletderived growth factor D,PDGFD)基因的结构及功能,并探究其在牦牛不同年龄的7个组织中的表达情况,本研究以36月龄大通牦牛的皮下脂肪组织cDNA为模板克隆牦牛PDGFD基因的CDS序列,利用基因序列进行生物信息学分析及蛋白理化性质预测,qPCR检测PDGFD在大通牦牛胚胎期150 d与36月龄成年时期2个年龄段7个组织(心,肝,脾,肺,肾,背最长肌,皮下脂肪)中的相对表达量。结果表明,大通牦牛PDGFD的CDS区全长为1095 bp,共编码氨基酸364个,同野牦牛PDGFD基因的亲缘关系最近(99.8%),与猪(Sus scrofa)最远(91.2%);牦牛PDGFD蛋白预测为稳定的亲水性分泌蛋白,无跨膜结构域,存在1个CUB结构域(116 aa)及1个PDGF结构域(93 aa);PDGFD蛋白高级结构由无规则卷曲(48.21%)、α-螺旋(25.62%)、延伸链(21.49%)和β-转角(4.68%)相互连接而成,主要与同为血小板源性生长因子、血管内皮生长因子相关蛋白相互作用。qPCR结果显示,牦牛2个发育阶段的7个组织中均有PDGFD基因的表达,在对比2个时期的PDGFD基因组织表达规律后发现,心脏中的基因表达维持在较高水平,脂肪组织中的基因表达量变化最大,随着牦牛生长极显著升高(P<0.001)。推测PDGFD基因与牦牛心脏发育有着密切关系,在生长发育过程中影响脂肪的沉积。本研究为PFGFD基因在牦牛脂肪沉积的调控机制的研究提供理论依据,可以作为牦牛的遗传育种的参考。Yak(Bos mutus)is an important genetic resource to the plateau region.To study the structure and function of the platelet-derived growth factor D(PDGFD)gene in yaks,and explore the PDGFD gene expression in different tissues at 2 age stages,this study cloned the complete PDGFD gene CDS sequence by using subcutaneous adipose tissue cDNA of 36-month-old Datong yak as the template.Biological software was used for analyzing the PDGFD gene nucleotide sequence and physical and chemical properties of protein.The rela-tive expression of PDGFD in 7 tissues(heart,liver,spleen,lungs,kidney,longissimus muscle and adipose tissue)of embryonic and 36-months old yak was carried out by qPCR.The results showed that the CDS region of PDGFD gene was 1095bp and encoded 364 amino acids.The most closely genetic relationship of Datong yak was the B.mutus(99.8%),and the most distant was the Sus scrofa(91.2%).The protein analysis of PDGFD showed that PDGFD contains 1 CUB domain of 116 aa,1 PDGF domain of 93 aa,and no transmembrane structure.PDGFD was predicted to be a stably hydrophilic secretory protein.The PDGFD protein advanced structure was consisted of random coil(48.21%),α-helix(25.62%),extended chain(21.49%),andβ-turn(4.68%).In addition,PDGFD mainly interacted with platelet-derived growth factor protein and vascular endothelial growth factor protein.qPCR results showed that PDGFD mRNA was expressed in each tissue at 2 stages.By comparing the 2 periods,PDGFD gene expression was maintained at a high level in the heart.Meanwhile,the expression in adipose tissue changed the most.The mRNA expression level of PDGFD extremely significantly increased in adipose tissue with the growth and development of yaks(P<0.001).The result showed that PDGFD may be closely related to yak's heart,and affected adipose growth and development.This study offers a theoretical foundation for the regulation mechanism of yak fat deposition and provides references for the genetic breeding of yaks.

关 键 词：大通牦牛 血小板源性生长因子D(PDGFD)基因 蛋白功能分析 脂肪组织 基因表达 

分 类 号：S929.9[农业科学—渔业资源]