斜带石斑鱼转化生长因子β1(TGF-β1)基因的克隆及表达分析

Molecular Cloning and Expression Analysis of Transforming Growth Factor-β1 (TGF-β1) from Orange-Spotted Grouper (Epinephelus coioides)

转化生长因子β1(TGF-β1)是一种高度保守的多效细胞因子。通过分子克隆得到了斜带石斑鱼Epi-nephelus coioidesTGF-β1 cDNA全长序列,全长为2 477 bp,开放阅读框为1 161 bp,编码一种含386个氨基酸的多肽。Realtim e PCR分析表明,TGF-β1基因在健康斜带石斑鱼所取组织中均有表达,肾脏中表达最高,其次是头肾和脾脏。用PolyI:C或ConA刺激斜带石斑鱼头肾淋巴细胞,TGF-β1表达量明显上升,且在刺激4 h后达到最高值。用PolyI:C刺激16、24 h后,TGF-β1几乎不表达,而用ConA刺激实验组,TGF-β1表达量一直高于对照组。结果说明,斜带石斑鱼TGF-β1基因结构和表达特征与其他硬骨鱼类具有高度相似性。

Transforming growth factor-β1（TGF-β1） is a highly conserved pleiotropic cytokine.In this study,we presented the bioinformatics analysis of TGF-β1 full-length cDNA sequences from orange-spotted grouper（Epinephelus coioides）.The full length cDNA is 2477 bp,which contains an ORF of 1161 bp,encoding a 386 amino acids protein.Real-time PCR results showed that TGF-β1 mRNA was predominantly expressed in the kidney,followed by head kidney and spleen of the healthy orange-spotted grouper.The expression of TGF-β1 in isolated headkidney lymphocytes was up-regulated treated with PolyI：C or ConA,and peaked at 4 hours after stimulation.Lymphocytes groups which were treated with PolyI：C had little TGF-β1 expression after 16 and 24 hours,while the expression of TGF-β1 was significantly higher than the control groups during the whole experimental processing time treated with ConA.In conclusion,we found the structure and expression pattern of grouper TGF-β1 was similar to those of other fish.