申玉建, 方程, 邱庆庆, 司景磊, 吴延军, 杨祝良, 杨秀荣, 郭亚芬, 蒋和生. 2017: 广西巴马小型猪PPARγ基因克隆及组织表达差异分析. 南方农业学报, 48(10): 1884-1890. DOI: 10.3969/j.issn.2095-1191.2017.10.24
引用本文: 申玉建, 方程, 邱庆庆, 司景磊, 吴延军, 杨祝良, 杨秀荣, 郭亚芬, 蒋和生. 2017: 广西巴马小型猪PPARγ基因克隆及组织表达差异分析. 南方农业学报, 48(10): 1884-1890. DOI: 10.3969/j.issn.2095-1191.2017.10.24
shu
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. 2017: Cloning of gene PPARγfrom Guangxi Bama mini-pig and tissue expression difference analysis for it. Journal of Southern Agriculture, 48(10): 1884-1890. DOI: 10.3969/j.issn.2095-1191.2017.10.24
Citation: 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. 2017: Cloning of gene PPARγfrom Guangxi Bama mini-pig and tissue expression difference analysis for it. Journal of Southern Agriculture, 48(10): 1884-1890. DOI: 10.3969/j.issn.2095-1191.2017.10.24
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广西巴马小型猪PPARγ基因克隆及组织表达差异分析

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

    摘要
  • 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GenBank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(pI)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.

     

    Abstract: ObjectiveIn 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 study-ing the mechanism of gene PPARγ in mycoplasmal pneumoniae of swine(MPS).MethodGene PPARγ from Guangxi Bama mini-pig was cloned by RT-PCR,and BLAST comparison analysis and bioinformatics analysis on its deduced pro-tein were conducted. The expressions of gene PPARγ were detected through real-time fluorescence quantitative PCR in heart,spleen,lung,kidney,large intestine,small intestine,muscle and fat of Guangxi Bama mini-pig of 18 months.ResultThe results showed that open reading frame of PPARγgene from Guangxi Bama mini-pig was 1515 bp in length, encoding 504 amino 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,reaching 100.0%;Guangxi Bama mini-pig shared 92.9%,91.8%,91.5%,90.5%and 83.5%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 PPARγ was highly conservative. The molecular weight of protein PPARγ was 57380.91 Da,its theoretical isoelectric point(pI)was 6.88,coefficient of instability was 49.26,fat index was 88.21,hydrophilic average was-0.293,which indicated that PPARγwas hydrophilic protein. The secondary structure was mainly alpha helix,accounting for 39.09%of the total. The expression of gene PPARγin large in-testine 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.ConclusionGene PPARγ expresses in all tissues of Guangxi Bama mini-pig of 18 months,but the expression shows significant difference in different tissues,which may be related to its functional differences in various tissues.

     

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