牦牛PAFR基因生物信息学分析及其在妊娠前后子宫中的表达

Cloning and Sequence Analysis of Yak PAFR Gene and Expression in Uterus Before and After Pregnancy

以牦牛血小板活化因子受体(Platelet-activating factor receptor,PAFR)基因为研究对象,采取西宁家养牦牛子宫为材料,采用RT-PCR技术克隆牦牛PAFR基因,对其进行生物信息学分析,并采用QRT-PCR方法定量检测PAFR在牦牛子宫不同时期的表达。结果表明,牦牛PAFR基因编码区长1 029 bp,编码342个氨基酸;氨基酸序列与黄牛同源性最高为99.7%,与非洲爪蟾蜍同源性最低为59.1%,表明PAFR氨基酸在进化过程中具有保守性;其编码蛋白的氨基酸序列有5个跨膜区域,推测其可能为跨膜蛋白;具有亲水性;未发现信号肽片段;含有G蛋白偶联受体家族结构域;QRT-PCR检测PAFR在牦牛子宫中表达,妊娠期最高,卵泡期最低,黄体期居中,揭示其与胚胎附植,妊娠维持有关。

Platelet-activating factor receptor（PAFR）gene were cloned from uterus of domestic yak by reverse transcription polymerase chain reaction（RT-PCR）. The results shows that domestic yak PAFR gene was 1 029 bp in length, encoded predicted mature protein of 342 amino acids, which encoded a hydrophilic protein. Compared with the reference sequence of PCR primer designing homology of the domestic yak PAFR coding sequence was 99.7%at the amino acid level, and also showed high homology of with other species. The PAFR has 5 transmembrane domains, features G protein-coupled receptor domains, no signal peptide. The result of the quantitative Real-time Polymerase chain reaction （QRT-PCR）shows that PAFR was highest expression in gestation and lowest in follicular phase in uterus of domestic yak. That suggested PAFR play an important role in implantation and gestation.