CAJ | 학술논문

用RACE-PCR和RT-PCR方法获得鳜(Siniperca chuatsi)膜结合型免疫球蛋白D(Membrane-bounded IgD,mIgD)重链基因的全长cDNA序列.鳜mIgD的cDNA全长为3 358 bp,其5'非编码区包含30 bp,3'非编码区包含337 bp;开放阅读框包含2991 bp,编码996个氨基酸,基因结构为VDJ-μ1-δ1-δ2-δ3-δ4-δ5-δ6-δ7-TM.鱼类IgD恒定区氨基酸序列比对结果显示,鳜mIgD存在半胱氨酸和色氨酸保守位点,与其他鱼类IgD的相似性在37%～72%之间.用邻接法(Neighbor Joining)构建的鱼类免疫球蛋白基因的系统发育树表明,鱼类IgD形成独立的一支,鳜mIgD与牙鲆和庸鲽IgD聚为一支.荧光定量PCR结果显示,鳜mIgD的mRNA主要分布于外周血白细胞、胸腺、头肾、中肾和脾脏中.
용RACE-PCR화RT-PCR방법획득궐(Siniperca chuatsi)막결합형면역구단백D(Membrane-bounded IgD,mIgD)중련기인적전장cDNA서렬.궐mIgD적cDNA전장위3 358 bp,기5'비편마구포함30 bp,3'비편마구포함337 bp;개방열독광포함2991 bp,편마996개안기산,기인결구위VDJ-μ1-δ1-δ2-δ3-δ4-δ5-δ6-δ7-TM.어류IgD항정구안기산서렬비대결과현시,궐mIgD존재반광안산화색안산보수위점,여기타어류IgD적상사성재37%～72%지간.용린접법(Neighbor Joining)구건적어류면역구단백기인적계통발육수표명,어류IgD형성독립적일지,궐mIgD여아평화용접IgD취위일지.형광정량PCR결과현시,궐mIgD적mRNA주요분포우외주혈백세포、흉선、두신、중신화비장중.
Full cDNA sequence of membrane-bounded immunoglobulin D(mIgD)heavy chain in the mandarin fish,Siniperca chuatsi has been isolated and characterized using RACE-PCR and RT-PCR methods.The mandarin fish' mlgD gene consists of 3 358 nucleotides including 5'UTR of 30 nueleotides,3' UTR of 337 nueleotides and an open reading frame with 2 991 nucleotides encoding 996 amino acid residues.The structure of mandarin fish' mIgD contained the VDJ region,the constant region of μ1-δ1-δ2-δ3-δ4-δ5-δ6-δ7,and TM region.The amino acid sequence of the constant region of mandarin fish mlgD shared 37%-72% identity with those of previously reported teleost IgDs including conserved cysteines and tryptophans.Phylogenetic tree based on teleost IgD heavy chain constant regions generated by Neighbor Joining method suggests that all teleosts cluster together,and that the mandarin fish' mIgD is clustered closely with Japanese flounder's IgD and Atlantic halibut's IgD.Real-time PCR showed that the mandarin fish' mIgD transcription was mainly detected in peripheral blood leucocytes(PBLs),thymus,head kidney,kidney and spleen.