畜牧兽医学报 ›› 2014, Vol. 45 ›› Issue (5): 699-705.doi: 10.11843/j.issn.0366-6964.2014.05.004

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GPR84和MPEG1基因的分子克隆和表达分析

王秋实1,2,武春艳1,2,李朋1,2,邵思宇1,2,张姣1,2,朱蒙1,2,杨秀芹1,2*   

  1. (1.黑龙江省普通高等学校动物遗传育种与繁殖重点实验室,哈尔滨 150030;2.东北农业大学动物科学技术学院,哈尔滨 150030)
  • 收稿日期:2013-11-13 出版日期:2014-05-23 发布日期:2014-05-23
  • 通讯作者: 杨秀芹,教授,博士生导师,E-mail:xiuqin163@163.com
  • 作者简介:王秋实(1987-),男,黑龙江双城人,硕士,主要从事动物遗传育种与繁殖研究,E-mail:wangqiushi87@163.com
  • 基金资助:

    国家自然科学基金项目(31072007)

Molecular Cloning and Expression Analysis of GPR84 and MPEG1 Genes in Porcine

WANG Qiu-shi1,2, WU Chun-yan1,2, LI Peng1,2, SHAO Si-yu1,2, ZHANG Jiao1,2, ZHU Meng1,2, YANG Xiu-qin1,2*   

  1. (1.Key Laboratory of Animal Genetics,Breeding and Reproduction,Education Department of Heilongjiang Province,Harbin 150030, China; 2.College of Animal Science and Technology,Northeast Agricultural University,Harbin 150030, China)
  • Received:2013-11-13 Online:2014-05-23 Published:2014-05-23
  • Supported by:
     

摘要:

本研究旨在克隆猪GPR84和MPEG1基因的完整编码区,对其进行序列分析,并研究它们的组织表达和诱导表达情况。以民猪为试验材料,应用RT-PCR方法克隆猪GPR84和MPEG1基因并进行生物信息学分析,利用实时荧光定量PCR方法构建组织表达谱,分析其在PK15细胞内的poly(I:C)诱导表达情况。结果表明,猪GPR84(GenBank accession No.JX280456)和MPEG1(GenBank accession No.JQ907392)基因编码区全长分别为1 191和2 154 bp,预期分别编码396和717个氨基酸残基的蛋白质,与人和牛相应蛋白序列的同源性均在83%以上。猪GPR84和MPEG1基因在11种组织中均有不同程度的表达,其中GPR84基因在肺中表达量最高,MPEG1基因在脾中表达量最高,二者在肌肉中的表达量均最低。在poly(I:C)的诱导下,GPR84基因的表达增强,MPEG1基因的表达则受到抑制。本研究成功地克隆了猪GPR84和MPEG1基因的全长编码区序列,并确定了它们的组织表达和诱导表达情况,为进一步揭示GPR84和MPEG1在抗病育种中的作用提供了基础。

Abstract:

The aim of this study was to clone the full-length coding sequence (CDS) of pig GPR84 and MPEG1 genes,and to characterize their expression profiles in tissues and induced by poly(I:C) in cultured PK-15 cells at mRNA level.The Minzhu was used in this study.The full-length CDSs of GPR84 and MPEG1 genes were cloned with RT-PCR method,and analyzed by using bioinformatic method.The expression profiles were analyzed using real-time quantitative RT-PCR method.The result showed that the full-length CDS of GPR84 (GenBank accession No.JX280456) and MPEG1 (GenBank accession No.JQ907392)genes were 1 191 and 2 154 bp,encoding 396 and 717 aa,respectively.Both of the proteins shared more than 83% similarities with their respective orthologous in human and cattle.The GPR84 and MPEG1 genes were expressed in all the eleven tissues analyzed with different abundance.The GPR84 gene was expressed at the most abundant level in lung,while the MPEG1 gene in spleen.Both were expressed at the least abundant level in muscle.Additionally,the expression of GPR84 was induced by poly(I:C) in PK15 cells,while MPEG1 was suppressed.The results will lay a foundation for further revealing the roles of GPR84 and MPEG1 in porcine disease-resistant breeding.

 

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