摘要应用同源克隆和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。
Abstract：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 .
收稿日期: 2014-12-25
引用本文:
李春涛,蒋自立,魏福伦,杨秀荣. 黄颡鱼免疫球蛋白3型轻链基因cDNA及其表达分析[J]. , 2014, (12): 814-819.
LI Chun-tao,JIANG Zi-Li,WEI Fu-lun,YANG Xiu-rong. cDNA Sequence and Expression Analysis of IgL3 Gene in Yellow Catfish, Pelteobagrus f ulvidraco. , 2014, (12): 814-819.
