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[目的]进行鲤鱼干扰素γ-2β(IFNγ-2β)全长cDNA的克隆、鉴定及序列分析。[方法]以鲤鱼外周血白细胞干扰素γ-2β(interferon γ-2β,IFNγ-2β)EST序列为基础,以地高辛标记作为探针对有丝分裂原刺激的鲤鱼外周血白细胞cDNA文库进行核酸杂交筛选,克隆鲤鱼IFNγ-2β的全长cDNA,并进行序列分析。[结果]获得了3个阳性克隆。对其序列分析结果显示,该序列包含119bp的5’非编码区,218bp的3’非编码区,开放阅读框ORF长537bp,共编码178个氨基酸,在其3’非编码区存在几个ATTTA不稳定基序。序列同源性比较结果表明,所获序列与GenBank上登录的鲤鱼IFN基因的同源性达97%;蛋白质序列和结构分析发现,该序列其具有IFN家族的典型序列特征。[结论]为进一步研究IFNγ-2β在体内的表达方式、功能特点和调控机理以及在炎症反应和免疫应答中的作用机制奠定了基础。
[Objective] The research aimed to clone, identify and sequence the full-length cDNA of IFNγ-2β. [Method] Based on the EST sequence of interferon γ-2β (IFNγ-2β) from carp, the cDNA library of mitogens stimulated carp peripheral blood leukocytes was digested with digoxigenin The full length cDNA of carp IFNγ-2β was cloned and sequenced. [Result] Three positive clones were obtained. The sequence analysis showed that the sequence contained 119bp 5 ’non-coding region and 218bp 3’ non-coding region. The open reading frame ORF was 537bp in length and encoded a total of 178 amino acids. There were several ATTTA in its 3 ’non-coding region Unstable motifs. The sequence homology comparison showed that the obtained sequence shared 97% homology with the common carp IFN gene registered in GenBank. The sequence and structure analysis of the sequence showed that the sequence shared the typical sequence features of IFN family. [Conclusion] This study lays the foundation for further study on the expression pattern, functional characteristics and regulatory mechanism of IFNγ-2β and the mechanism of action in inflammatory response and immune response.