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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(inter-feronγ-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,对其进行序列分析,结果显示,该序列包含119 bp的5’非编码区,218 bp的3’非编码区,开放阅读框ORF长537 bp,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 inter-feronγ-2β (IFNγ-2β) of carp, the cDNA library of mitogens stimulated carp peripheral blood leukocytes was probed with digoxigenin The full-length cDNA of IFNγ-2β was cloned by nucleic acid hybridization and sequenced. [Result] Three positive clones were obtained and sequenced. The results showed that the sequence contained 119 bp 5 ’non - coding region and 218 bp 3’ non - coding region. The open reading frame ORF was 537 bp in length, encoding a total of 178 amino acids. There are several ATTTA instability motifs in its 3 ’non-coding region. The sequence homology comparison shows that the sequence has 97% homology with the common carp IFN gene registered in GenBank. The protein sequence and Structural analysis revealed that this sequence has the typical sequence characteristics of the 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.