• 中文核心期刊
  • CSCD来源期刊
  • 中国科技核心期刊
  • CA、CABI、ZR收录期刊

闽西南黑兔NPFFR2基因的克隆与序列分析

Cloning and Bioinformatics of NPPFR2 of Minxinan Black Rabbit

  • 摘要:
      目的  神经肽FF受体2(Neuropeptide FF receptor 2,NPFFR2)是机体重要的神经内分泌因子神经肽FF(Neuropeptide FF,NPFF)的受体,参与糖皮质激素和焦虑行为的调节。测序分析闽西南黑兔NPFFR2基因,为其作为闽西南黑兔抗应激研究的重要靶基因提供数据。
      方法  克隆NPFFR2基因,并对其进行测序及序列分析。
      结果  成功克隆得到闽西南黑兔NPFFR2基因2个转录异构体,分别含1 815和1 824个核苷酸。它们拥有相同的5′-UTR(Untranslated region,非翻译区)和编码区序列(Coding region),但是在3′-UTR存在多个突变。根据mRNA二级结构预测结果,3′-UTR的突变致使NPFFR2基因转录异构体的二级结构存在部分差异。通过对兔NPFFR2基因进行遗传进化分析,发现其和人类的相关基因进化关系最近。依据编码序列对NPFFR2蛋白结构和亚细胞分布进行预测,NPFFR2蛋白含414个氨基酸,有7个跨膜α螺旋,是G蛋白偶联受体家族一员。
      结论  分离克隆了闽西南黑兔NPFFR2基因的2种转录异构体,并利用生物信息学方法对其转录本和翻译蛋白的结构和功能进行了预测,为进一步高效利用闽西南黑兔NPFFR2基因提供相关理论支持。

     

    Abstract:
      Objective  The gene related to an important neuroendocrine receptor of NPFF (neuropeptide FF), NPFFR2 (Neuropeptide FF receptor 2), that participates in the regulation of corticotropin-releasing hormone (CRT) and anxiety-like behaviors was analyzed for study on the stress response of Minxinan black rabbit.
      Method  NPFFR2 was cloned, sequenced, and analyzed.
      Result  Two transcript variants of NPFFR2 from Minxinan black rabbits were cloned to show the contents of 1 815 bp and 1 824 bp nucleic acid sequences. They had same sequences in 5′-UTR and coding region but multiple mutations in 3′-UTR. The predicted mRNA secondary structure displayed multiple mutations in the 3′-UTR which could cause changes on the structure as well as the function of NPFFR2. The phylogenetic analysis indicated the NPFFR2 from the rabbit was most closely related to that from Homo sapiens. According to the predicted protein structure and subcellular location, NPFFR2 protein belonged to the G protein-coupled receptors family and had 414 amino acids and 7 transmembrane α-helixes.
      Conclusion  Two transcript variants of NPFFR2 from Minxinan black rabbits were cloned. The structures and functions of the transcripts and translated proteins were predicted by bioinformatics obtained. The results would be helpful for further study on the functions of NPFFR2 of Minxinan black rabbit.

     

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