畜牧兽医学报 ›› 2010, Vol. 41 ›› Issue (9): 1068-1075.doi:

• 遗传繁育 • 上一篇    下一篇

猪PGC-1β和PRC基因的部分cDNA克隆及表达分析

常志光,卢荣华,吉红,苏尚顺,高晓娟,杨公社*   

  1. 西北农林科技大学动物科技学院 动物脂肪沉积与肌肉发育实验室,杨凌 712100
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2010-09-20 发布日期:2010-09-20
  • 通讯作者: 杨公社

Cloning and Expressional Analysis of PGC-1β and PRC cDNA in Pig

CHANG Zhi-guang, LU Rong-hua, JI Hong,SU Shang-shun,GAO Xiao-juan,YANG Gong-she*   

  1. Laboratory of Animal Fat Deposition and Muscle Development, College of Animal Science and Technology, Northwest A&F University, Yangling 712100, China
  • Received:1900-01-01 Revised:1900-01-01 Online:2010-09-20 Published:2010-09-20

摘要: 以猪心脏组织为材料,克隆猪PGC-1β(PPARγcoactivator-1β)和PRC(PGC-1-related coactivator)基因的cDNA序列,并进行生物信息学分析,同时明确其组织表达特征。根据人PGC-1β和PRC基因的已知mRNA序列和在猪ESTs数据库中搜索得到的同源序列,利用RT-PCR技术克隆猪PGC-1β和PRC基因的部分cDNA序列,并用SQ-RT-PCR和Western blot方法检测其在猪心脏、肌肉、脾脏、肝脏、肺脏、肾脏、小肠、脂肪组织中的表达。结果,获得长度分别为1 251和1 254 bp的猪PGC-1β和PRC基因的部分序列,分别编码415和417个氨基酸(GenBank登录号分别为:FJ377680和FJ 479791),其核苷酸序列和氨基酸序列与其它物种的同源性均达到80%左右,并包含有TPPTTPP和DHDY 2个保守性序列,以及RNA识别基序(RRM)的保守性结构域。PGC-1β在8个组织中均有表达,而PRC主要在心脏、肝脏、肾脏和骨骼肌中表达,在脾脏和小肠中未检测到。本研究成功获取了猪PGC-1β和PRC基因部分序列,且组织表达存在差异,所获得的结果为研究PGC-1家族的生理功能及调节机制积累了资料。

Abstract: The aim of this study was to clone the cDNA of PGC-1β and PRC gene from the heart of porcine by RT-PCR approach and to analyze its sequence and expression profile. According to the PGC-1β and PRC mRNA sequences of human which were the homologous sequences of pig searched in the EST library, the partial PGC-1β and PRC cDNA sequence of pigs were cloned by RT-PCR, and their expression profile were detected in heart, muscle, spleen, liver, lung, kidney, small intestine and adipose tissue by using RT-PCR and Western blot. The result showed that the PGC-1β and PRC were 1 251 and 1 254 bp in length, and encoding 415 and 417 amino acids(GenBank accession number:FJ377680 and FJ479491), respectively. Bioinformatic analysis found that the homology of nucleotide sequence and amino acid were about 80% compared with the counterpart sequences of other species, and containing TPPTTPP and DHDY conserved sequences, as well as the RNA recognition motif(RRM)conserved domains. PGC-1β was widely expressed in 8 tissues,while PRC was highly expressed only in heart, liver, skeletal muscle and kidney,but not detected in spleen and small intestine. The result suggested that the differention of expression level of the PGC-1β and PRC may be related to their physiological functions in different tissues.The results will be helpful for elucidating the physiological function and regulatory mechanisms of PGC-1 family.