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1251和1254bp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FJ479791),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TPPT-TPP和DHDY2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。
The aim of this study was to clone the cDNA of PGC-1β and PRCgene from the heart of porcine by RT-PCR approach and to analyze its sequence and expression profile.According to the PGC-1βand PRC mRNA sequences of human which were the homologous sequences of pig searched in the EST library,the partial PGC-1βand 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-1βand PRC were 1 251and 1 254bp in length,and encoding 415and 417amino acids（GenBank accession number：FJ377680and 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-1βwas widely expressed in 8tissues, while PRCwas 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 PRCmay be related to their physiological functions in different tissues.The results will be helpful for elucidating the physiological function and regulatory mechanisms of PGC-1family.
出处
《畜牧兽医学报》
CAS
CSCD
北大核心
2010年第9期1068-1075,共8页
ACTA VETERINARIA ET ZOOTECHNICA SINICA
基金
国家高技术研究发展计划(863计划)(2006AA10Z138)
中国博士后科学基金项目(20080431254)
