从江香猪λ1干扰素基因的克隆及序列分析

Cloning and bioinformatics analysis of Congjiang xiang pig IFN-λ1 gene

[目的]对从江香猪λ1干扰素(interferon-λ1,IFN-λ1)基因进行扩增、克隆和生物信息学分析。[方法]根据Gen Bank登录的猪IFN-λ1基因序列(FJ455508)设计合成特异性引物,通过RT-PCR从淋巴细胞中扩增IFN-λ1基因CDS区并进行克隆和生物信息分析。[结果]从江香猪IFN-λ1基因CDS全长576 bp,共编码191个氨基酸,其分子式为C949H1543N277O270S7,相对分子质量21.38 k Da;该蛋白等电点为9.42,为亲水性蛋白。从江香猪IFN-λ1含有丰富的二级结构,以α-螺旋(64.40%)和无规卷曲(25.65%)为主,蛋白质三级结构中主要以α-螺旋为主,同源性及系统进化树分析结果表明从江香猪与野猪IFN-λ1核苷酸同源性最高(为100%),亲缘关系最近。[结论]从江香猪IFN-λ1基因的克隆和生物信息学分析为进一步研究其抗病毒功能奠定了基础。

[Objective]To clone and proceed the biological information analysis of Congjiang xiang pig IFN-λ1 gene.[Methods]A pair of specific primers were designed to amplify the CDS region of Congjiang xiang pig IFN-λ1 gene. The IFN-λ1 gene was amplified from lymphocyte cells by RT-PCR and then analyzed by bioinformatics tools. [Results] The full-length CDS region of Congjiang xiang pig IFN-λ1 gene was 576 bp and encoded 191 amino acids. The molecular formula of IFN-λ1 protein was C949 H1543 N277 O270 S7 with a relative molecular mass about 21. 38 k Da and p I 9. 42. Protein secondary structure prediction showed that Congjiang xiang pig IFN-λ1 protein contained abundant secondary structure and were mainly alpha helix（64. 90%） and random coil（25. 65%）. Protein tertiary structure mainly contained alpha helix. The results of gene homology and phylogenetic tree revealed that Congjiang xiang pig IFN-λ1 gene had the closest relationship with sus scrof.[Conclusion]The cloning and bioinformatics analysis of Congjiang xiang pig IFN-λ1 gene provided the theoretical foundation for further studying its functions.