水产科学
水產科學
수산과학
FISHERIES SCIENCE
2014年
12期
814-819
,共6页
李春涛%蒋自立%魏福伦%杨秀荣
李春濤%蔣自立%魏福倫%楊秀榮
리춘도%장자립%위복륜%양수영
黄颡鱼%IgL3%cDNA%实时 PCR%表达
黃顙魚%IgL3%cDNA%實時 PCR%錶達
황상어%IgL3%cDNA%실시 PCR%표체
Pelteobagrus f ulvidraco%IgL3%cDNA%Real-time PCR%expression
应用同源克隆和RACE‐PCR方法获得黄颡鱼免疫球蛋白3型轻链(IgL3)基因全长cDNA ,并分析了该基因在组织中的表达。黄颡鱼IgL 3的cDNA全长为965 bp ,包含5′53 bp非编码区,3′189 bp非编码区,开放阅读框723 bp ,编码241个氨基酸。黄颡鱼与其他6种硬骨鱼类Ig L 3氨基酸序列比对分析表明,黄颡鱼Ig L 3氨基酸序列与大鳍鱯Ig L 3型的相似性最高,为72.8%,与大西洋鲑Ig L 3型的相似性最低,为42.2%。进化树分析表明,黄颡鱼Ig L与大鳍鱯Ig L 3型聚为一支同时与其他鱼类IgL 3型聚为一簇,明显与L1和L2进化支不同。实时PCR显示,黄颡鱼IgL3基因主要在头肾、脾脏、血细胞、鳃和肠中转录表达;注射嗜水气单胞菌后,头肾、脾脏和血细胞IgL3基因表达量有显著上升。表明这些组织是黄颡鱼Ig L 3型基因主要的表达器官,在免疫反应具有重要作用。
應用同源剋隆和RACE‐PCR方法穫得黃顙魚免疫毬蛋白3型輕鏈(IgL3)基因全長cDNA ,併分析瞭該基因在組織中的錶達。黃顙魚IgL 3的cDNA全長為965 bp ,包含5′53 bp非編碼區,3′189 bp非編碼區,開放閱讀框723 bp ,編碼241箇氨基痠。黃顙魚與其他6種硬骨魚類Ig L 3氨基痠序列比對分析錶明,黃顙魚Ig L 3氨基痠序列與大鰭鱯Ig L 3型的相似性最高,為72.8%,與大西洋鮭Ig L 3型的相似性最低,為42.2%。進化樹分析錶明,黃顙魚Ig L與大鰭鱯Ig L 3型聚為一支同時與其他魚類IgL 3型聚為一簇,明顯與L1和L2進化支不同。實時PCR顯示,黃顙魚IgL3基因主要在頭腎、脾髒、血細胞、鰓和腸中轉錄錶達;註射嗜水氣單胞菌後,頭腎、脾髒和血細胞IgL3基因錶達量有顯著上升。錶明這些組織是黃顙魚Ig L 3型基因主要的錶達器官,在免疫反應具有重要作用。
응용동원극륭화RACE‐PCR방법획득황상어면역구단백3형경련(IgL3)기인전장cDNA ,병분석료해기인재조직중적표체。황상어IgL 3적cDNA전장위965 bp ,포함5′53 bp비편마구,3′189 bp비편마구,개방열독광723 bp ,편마241개안기산。황상어여기타6충경골어류Ig L 3안기산서렬비대분석표명,황상어Ig L 3안기산서렬여대기화Ig L 3형적상사성최고,위72.8%,여대서양해Ig L 3형적상사성최저,위42.2%。진화수분석표명,황상어Ig L여대기화Ig L 3형취위일지동시여기타어류IgL 3형취위일족,명현여L1화L2진화지불동。실시PCR현시,황상어IgL3기인주요재두신、비장、혈세포、새화장중전록표체;주사기수기단포균후,두신、비장화혈세포IgL3기인표체량유현저상승。표명저사조직시황상어Ig L 3형기인주요적표체기관,재면역반응구유중요작용。
The technique of homologous cloning and RACE was used to amplify full length cDNA of IgL 3 gene from yellow catfish (Pelteobagrus f ulvidraco) .IgL3 in yellow catfish has 965 nucleotides ,including 5′‐UTR of 53 nucleotides ,3′‐UTR of 189 nucleotides and an open reading frame with 723 nucleotides en‐coding a peptide of 241 amino acids .The IgL3 comparison in seven teleost species showed that IgL 3 in yellow catfish shared the maximal identity (72 .8% ) with that in Mystus macropterus ,and the minimal i‐dentity(42 .2% ) with that in Atlantic salmon Salmo salar .Phylogenetic tree based on some teleost IgL a‐mino acids showed that IgL3 in yellow catfish was clustered closely with that of M .macropterus and IgL3 was far away from L1 and L2 of teleost .Real‐time PCR revealed that IgL3 mRNA expression of yellow catfish was mainly detected in head kidney ,spleen ,blood cells ,gill and intestine and increased significant‐ly in these tissues after injection of Aeromonas hydrophila .The results indicated that these tissues were main responding organs for IgL3 expression after stimulation ,and played a critical role in immunity inter‐action in yellow catfish .