CAJ | 학술논문

根据GenBank中登录的仿刺参Apostichopus japonicus ghitm基因的EST片段,采用RACE扩增法克隆了仿刺参ghitm基因的cDNA全长序列,并利用qRT-PCR技术检测了经LPS诱导后仿刺参ghitm的表达变化情况。结果表明：仿刺参ghitm基因( Aj-ghitm)的cDNA序列全长为1325 bp,包含一个1005 bp编码334个氨基酸的开放阅读框,序列两端分别为140 bp的5’UTR和180 bp的3’UTR。序列分析结果显示, Aj-ghitm编码的蛋白含有一个8次跨膜结构域,与GenBank中登录的紫色球海胆GHITM蛋白的氨基酸序列相似度为65·4%,且在系统发育树中聚为一支； qRT-PCR检测结果表明,经LPS刺激可诱导仿刺参体腔细胞Aj-ghitm mRNA的表达且呈先下降后升高最后回复到接近正常水平的趋势,在24 h时上升到最高,随后下降。本研究是有关棘皮动物ghitm基因的首次报道,其结果可为进一步探讨ghitm在仿刺参抗细菌感染以及生长发育等方面的功能及作用机制提供参考。
근거GenBank중등록적방자삼Apostichopus japonicus ghitm기인적EST편단,채용RACE확증법극륭료방자삼ghitm기인적cDNA전장서렬,병이용qRT-PCR기술검측료경LPS유도후방자삼ghitm적표체변화정황。결과표명：방자삼ghitm기인( Aj-ghitm)적cDNA서렬전장위1325 bp,포함일개1005 bp편마334개안기산적개방열독광,서렬량단분별위140 bp적5’UTR화180 bp적3’UTR。서렬분석결과현시, Aj-ghitm편마적단백함유일개8차과막결구역,여GenBank중등록적자색구해담GHITM단백적안기산서렬상사도위65·4%,차재계통발육수중취위일지； qRT-PCR검측결과표명,경LPS자격가유도방자삼체강세포Aj-ghitm mRNA적표체차정선하강후승고최후회복도접근정상수평적추세,재24 h시상승도최고,수후하강。본연구시유관극피동물ghitm기인적수차보도,기결과가위진일보탐토ghitm재방자삼항세균감염이급생장발육등방면적공능급작용궤제제공삼고。
The full-length cDNA sequence of ghitm were firstly cloned by RACE technique from coelomocytes of sea cucumber Apostichopus japonicus according to the EST sequence previously obtained from Genbank, and we named it as Aj-ghitm ( GenBank:KC886718 ) . The full-length cDNA of Aj-ghitm was 1325 bp, containing a 5’UTR of 140 bp and a 3’UTR of 180 bp, with an open reading frame of 1005 bp encoding 334 amino acids.SMART analysis showed that the deduced protein has an eight-transmembrane domain. Multiple alignment and phylogenic analysis showed that Aj-GHITM and the sea urchin GHITM were closely clustered in one group with the highest similarity of 65 . 4%. Quantitative real-time PCR revealed that Aj-ghitm mRNA expression level was de-creased first and then increased, finally recovery to the normal level after LPS-induced, with the maximum expres-sion 24 h followed by a lower expression. The findings give a reference for further study of the role of GHITM in the development and anti-bacterial infection in sea cucumber.