CAJ | 학술논문

[目的]进行鲤鱼白细胞介素-1β(IL-1β)全长cDNA的克隆、鉴定及其差异表达分析.[方法]利用DD-RTPCR方法获得差异表达cDNA片段,对有丝分裂原刺激的鲤鱼外周血白细胞cDNA文库进行筛选,克隆了鲤鱼IL-1β的全长cDNA,并进行了序列分析和差异表达分析.[结果]获得的阳性克隆含有1个大小为831 bp编码276个氨基酸的完整开放阅读框.聚类分析表明,鲤鱼IL-1β氨基酸序列与日本鲤鱼紧密聚为一支,氨基酸序列的同源性达95%,之后聚类顺序依次为鲫鱼、斑马鱼、猪、牛、马、人和小鼠.差异表达分析表明,经有丝分裂原刺激后前期(2 h)白细胞中IL-1β的表达量显著增大,但随着时间推移(12,24 h)并非一直较同期大,表达量总体趋势成峰形.[结论]为进一步研究IL-1β在体内的表达方式、功能特点和调控机理以及在炎症反应、应急反应和免疫应答的作用机制奠定了基础.
[목적]진행리어백세포개소-1β(IL-1β)전장cDNA적극륭、감정급기차이표체분석.[방법]이용DD-RTPCR방법획득차이표체cDNA편단,대유사분렬원자격적리어외주혈백세포cDNA문고진행사선,극륭료리어IL-1β적전장cDNA,병진행료서렬분석화차이표체분석.[결과]획득적양성극륭함유1개대소위831 bp편마276개안기산적완정개방열독광.취류분석표명,리어IL-1β안기산서렬여일본리어긴밀취위일지,안기산서렬적동원성체95%,지후취류순서의차위즉어、반마어、저、우、마、인화소서.차이표체분석표명,경유사분렬원자격후전기(2 h)백세포중IL-1β적표체량현저증대,단수착시간추이(12,24 h)병비일직교동기대,표체량총체추세성봉형.[결론]위진일보연구IL-1β재체내적표체방식、공능특점화조공궤리이급재염증반응、응급반응화면역응답적작용궤제전정료기출.
[Obiective] The research aimed to carry out the cloning, identification and differential expression analysis of carp interleukin-1β(1L-1β) cDNA. [Method] By using DD-RTPCR method, the differential expression cDNA fragments were gained. The cDNA library of carp peripheral blood leucocytes which was stimulated by the mitogen was screened, and the full length cDNA of carp IL-1β was cloned. Moreover, the sequence analysis and differential expression analysis were carried out. [Result] The positive clone which had a whole ORF that encoded 276 amino acids was obtained. The cluster analysis showed that the amino acid sequence of carp IL-1 β and Japanese carp closely gathered as a branch, and the homoeology of amino acid sequence reached 95%. The clustering order was the carassius, zebra fish, pig, cattle, horse, human and mouse in turn. The differential expression analysis showed that the expression of IL-1 β in the leucocytes significantly increased in the prior period (4 h) after the mitogen stimulated. But as the time went by (12 and 24 h ), it didn't increase in the same period. The total trend of expression amount presented the peak type. [Conclusion] The research laid the foundation for further studying the expression manner, function characteristic, regulation mechanism of IL-1β in vivo and its action mechanisms in the inflammatory reaction, emergency reaction and immune response.