丙型肝炎病毒非结构蛋白5A结合蛋白37小鼠同源基因的克隆化及结构分析

Identification and sequence analysis of mouse homologous gene coding for hepatitis C virus non-structural protein 5A-binding protein 37

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 murine homologous gene to human NS5ABP37 gene, and to elucidate its biological functions and the possible effects in the pathogenesis of hepatitis C virus (HCV) infection. METHODS: Yeast-two hybrid system was employed to screen the human liver cDNA library by using non-structural protein 5A (NS5A) of hepatitis C virus (HCV) as the bait. According to the homologous role between the species, a murine NS5ABP37 homologous to human NS5ABP37 was deduced by bioinformatics methods. The primary sequence of murine NS5ABP37 was searched for conserved domains by online tools of GeneBank. RESULTS: Human NS5ABP37 was screened and cloned from human liver cDNA library by yeast-two hybrid system 3. The murine NS5ABP37 was deduced by bioinformatics methods. The open reading frame (ORF) of murine NS5ABP37 con- sisted of 1 488 nt encoding a protein of 495 amino acids. Calculated molecular weight was 54 583.67 dalton and predicted pI was 4.70. From the homologous protein search, murine NS5ABP37 was demonstrated homologous to leukocyte antigen related (LAR) protein precursor. The sequence of murine NS5ABP37 was deposited into GenBank, and the accession number was AY234860. CONCLUSION: Successful identification and cloning of murine NS5ABP37, which is homologous to LAR protein precursor, paves a way for elucidating the biological function of murine NS5ABP37 and pathogenesis of hepatitis C virus infection.