广西巴马小型猪PPARγ基因克隆及组织表达差异分析  

Cloning of gene PPARγ from Guangxi Bama mini-pig and tissue expression difference analysis for it

作　　者：申玉建 方程[1] 邱庆庆 司景磊 吴延军[1] 杨祝良[1] 杨秀荣[1] 郭亚芬[1] 蒋和生[1] SHEN Yu-jian;FANG Cheng;QIU Qing-qing;SI Jing-lei;WU Yan-jun;YANG Zhu-liang;YANG Xiu-rong;GUO Ya-fen;JIANG He-sheng(College of Animal Science & Technology,Guangxi University,Nanning 530004,China)

机构地区：[1]广西大学动物科学技术学院,南宁530004

出　　处：《南方农业学报》2017年第10期1884-1890,共7页Journal of Southern Agriculture

基　　金：国家现代农业产业技术体系广西生猪创新团队建设项目(nycytxgxcxtd-03-15);广西家畜遗传育种改良重点实验室开放项目(2015GXKLLGI-09)

摘　　要：【目的】克隆广西巴马小型猪过氧化物酶增殖物激活受体γ基因(PPARγ),并确定其在不同组织中的表达情况,为后续研究该基因在猪支原体肺炎(MPS)中的作用机制打下基础。【方法】利用RT-PCR克隆广西巴马小型猪PPARγ基因,并进行BLAST比对分析及其推导蛋白的生物信息学分析;以实时荧光定量PCR检测PPARγ基因在18月龄广西巴马小型猪心脏、脾脏、肺脏、肾脏、大肠、小肠、肌肉和脂肪中的表达情况。【结果】广西巴马小型猪PPARγ基因开放阅读框序列1515 bp,编码504个氨基酸。广西巴马小型猪PPARγ基因推导氨基酸序列与Gen Bank已发表的猪PPARγ基因(FJ436399.1)推导氨基酸序列同源性最高,达100.0%;而与牛(NM_181024.2)、人类(NM_005037.5)、猕猴(NM_001032860.1)、鼠(NM_001308354.1)和鸿雁(KJ019822.1)的同源性分别为92.9%、91.8%、91.5%、90.5%和83.5%,表明PPARγ基因高度保守。PPARγ蛋白分子量为57380.91 Da,理论等电点(p I)为6.88,不稳定系数49.26,脂肪指数为88.21,亲水性平均水平为-0.293,为亲水性蛋白,其二级结构主要是α-螺旋,占总数的39.09%。PPARγ基因在广西巴马小型猪大肠组织中的表达量最高,极显著高于其他组织(P<0.01),而在肾脏、心脏和肌肉组织中的表达量极低。【结论】PPARγ基因在18月龄广西巴马小型猪的各组织中均有表达,但不同组织中的表达水平存在明显差异,可能与其在不同组织中的功能差异有关。weight of protein PPARγwas57380.91Da,its theoretical isoelectric point(pI)was6.88,coefficient of instability was49.26,fat index was88.21,hydrophilic average was-0.293,which indicated that PPARγwas hydrophilic protein.The secondary structure was mainly alpha helix,accounting for39.09%of the total.The expression of gene PPARγin large intestine of Guangxi Bama mini-pig was the highest,extremely significantly higher than those in other tissues(P<0.01),while its expression was very low in kidney,heart and muscle.【Conclusion】Gene PPARγexpresses in all tissues of Guangxi Bama mini-pig of18months,but the expression shows significant difference in different tissues,which may be related to its functional differences in various tissues.【Objective】In this study,peroxisome proliferator-activated receptorγgene(PPARγ)from Guangxi Bama mini-pig was cloned and its expressions in different tissues were determined in order to lay a foundation for further studying the mechanism of gene PPARγin mycoplasmal pneumoniae of swine(MPS).【Method】Gene PPARγfrom Guangxi Bama mini-pig was cloned by RT-PCR,and BLAST comparison analysis and bioinformatics analysis on its deduced protein were conducted.The expressions of gene PPARγwere detected through real-time fluorescence quantitative PCR in heart,spleen,lung,kidney,large intestine,small intestine,muscle and fat of Guangxi Bama mini-pig of18months.【Result】The results showed that open reading frame of PPARγgene from Guangxi Bama mini-pig was1515bp in length,encoding504amino acids.The homology between the deduced amino acid sequence of gene PPARγfrom Guangxi Bama mini-pig and the published porcine gene PPARγ(FJ436399.1)in GenBank were the highest,reaching100.0%;Guangxi Bama mini-pig shared92.9%,91.8%,91.5%,90.5%and83.5%homology with amino acid sequences from Bos taurus(NM_181024.2),Homo sapiens(NM_005037.5),Macaca mulatta(NM_001032860.1),Mus musculus(NM_001308354.1),and Anser cygnoides(KJ019822.1)respectively,which indicated that PPARγwas highly conservative.The mole

关 键 词：广西巴马小型猪 PPARΓ基因 猪肺炎支原体 克隆 组织表达差异 炎症调控 

分 类 号：S828.89[农业科学—畜牧学]