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目的:克隆丙型肝炎病毒(HCV)非结构蛋白5A(NS5A)结合蛋白37(NS5ABP37)的小鼠同源基因,为阐明小鼠NS5ABP37基因的生物学功能奠定基础,探索HCVNS5A蛋白与NS5ABP37蛋白之间的结合在慢性丙型肝炎的发病机制中可能的作用.方法:利用酵母双杂交系统3,以HCVNS5A蛋白作为诱饵,筛选肝细胞cDNA文库,首先获得HCVNS5A结合的人肝细胞蛋白的新的编码基因.利用生物信息学(bioinformatics)技术确定NS5ABP37的小鼠同源基因序列.利用GenBank在线分析软件,对小鼠NS5ABP37蛋白一级结构特点进行分析.结果:通过酵母双杂交技术获得了人NS5ABP37的编码基因.利用生物信息学技术确定了小鼠NS5ABP37的基因序列.利用分子生物学技术获得了小鼠的NS5ABP37的基因克隆,小鼠NSSABP37基因开放读码框架(ORF)为1488nt,编码产物由495aa组成.计算分子量为54583.67道尔顿,预测等电点(pI)为4.70.小鼠NS5ABP37与白细胞抗原相关(leukocyteantigenrelated,EAR)蛋白前体蛋白(LARproteinprecursor)同源.小鼠HCVNS5ABP37基因序列被美国核苷酸数据库GenBank收录,收录号为AY234860.结论:成功确定、克隆了小鼠的NS5ABP37基因序列,并证实与白细胞抗原相关蛋白前体蛋白具有一定的同源性.这一结果,为进一步阐明小鼠的NS5ABP37基因的功能,阐明HCV感染的发病机制奠定了坚实的基础.
AIM: To clone the mouse homologous gene of hepatitis C virus (NS5A) binding protein 37 (NS5ABP37) and lay a foundation for elucidating the biological function of mouse NS5ABP37 gene and to explore the relationship between HCV NS5A protein and NS5ABP37 protein In the pathogenesis of chronic hepatitis C.Methods: The yeast two-hybrid system was used to screen the hepatocyte cDNA library with HCVNS5A protein as a decoy, and then the new coding of HCVNS5A-bound human hepatocyte protein was obtained The bioinformatics technique was used to identify the mouse NS5ABP37 homologous gene sequence.The GenBank online analysis software was used to analyze the primary structure of mouse NS5ABP37 protein.Results: The NS5ABP37 The gene sequence of mouse NS5ABP37 was determined by bioinformatics technique.The mouse NS5ABP37 gene was cloned by molecular biology technique.The mouse NSSABP37 gene open reading frame (ORF) was 1488nt and the encoded product was composed of 495aa The calculated molecular weight was 54583.67 Daltons with a predicted isoelectric point (pI) of 4.70. The mouse NS5ABP37 was associated with leukocyteant igenrelated, EAR) protein precursor protein (LARproteinprecursor) homologous.The mouse HCVNS5ABP37 gene sequence was collected by the US nucleotide database GenBank, accession number AY234860.Conclusion: The mouse NS5ABP37 gene sequence was successfully confirmed and confirmed with Leukocyte antigen-related protein precursor protein has a certain degree of homology.This result provides a solid foundation for elucidating the function of mouse NS5ABP37 gene and elucidating the pathogenesis of HCV infection.