CAJ | 학술논문

[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。
[목적]진행리어간우소γ-2β（IFNγ-2β）전장cDNA적극륭、감정급서렬분석。[방법]이리어외주혈백세포간우소γ-2β（interferon γ-2β,IFNγ-2β）EST서렬위기출,이지고신표기작위탐침대유사분렬원자격적리어외주혈백세포cDNA문고진행핵산잡교사선,극륭리어IFNγ-2β적전장cDNA,병진행서렬분석。[결과]획득료3개양성극륭。대기서렬분석결과현시,해서렬포함119bp적5＇비편마구,218bp적3’비편마구,개방열독광ORF장537bp,공편마178개안기산,재기3’비편마구존재궤개ATTTA불은정기서。서렬동원성비교결과표명,소획서렬여GenBank상등록적리어IFN기인적동원성체97%;단백질서렬화결구분석발현,해서렬기구유IFN가족적전형서렬특정。[결론]위진일보연구IFNγ-2β재체내적표체방식、공능특점화조공궤리이급재염증반응화면역응답중적작용궤제전정료기출。
[Objective] The research aimed to carry out the cloning, identification and sequence analysis of full-length cDNA of carp interferon γ-2β （IFNγ-2β）. [Method] The cDNA library of peripheral blood leucocytes which were separated from carp and stimulated with mitogen was screened by a probe labeled with DIG. The IFNγ- 2β EST sequence was picked out from the constructed cDNA library of peripheral blood leucocyte, and the full length of carp interferon γ-2β was cloned. In addition, the sequence analysis was carried out. [Result] Three positive clones were obtained. Sequence analysis indicated that the sequence had a 119 bp 5’-UTR and a 218 bp 3’-UTR, and the open reading frame （ORF）of this gene was 537 bp which putatively coded 178 amino acids and there were several instable motifs for mRNA （ATTTA） in the 3’-untranslated region. Its homology with IFN from GenBank was up to 97% . Analysis on protein sequence and structure showed that the predicted protein sequence was identified as an IFN family signature. [Conclusion] The research laid the foundation for further studying the expression manner, function characteristic and regulation mechanism of IFNγ-2β in vivo and the action mechanism in the inflammatory reaction, emergency reaction and immune response.