中山大学学报自然科学版 ›› 2011, Vol. 50 ›› Issue (1): 92-98.

• 研究论文 • 上一篇    下一篇

斜带石斑鱼转化生长因子β1(TGFβ1)基因的克隆及表达分析

平海林,吴金英,徐胜威,胡开顺,段志刚   

  1. (中山大学生命科学学院水生经济动物研究所∥广东省水生经济动物良种繁育重点实验室,广东 广州 510275)
  • 收稿日期:2010-01-20 修回日期:1900-01-01 出版日期:2011-01-25 发布日期:2011-01-25
  • 通讯作者: 吴金英

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

PING Hailin,WU Jinying,XU Shengwei,HU Kaishun,DUAN Zhigang   

  1. (Institute of Aquatic Economic Animals and Guangdong Provincial Key Laboratory for Aquatic Economic Animals, School of Life Sciences, Sun Yatsen University, Guangzhou 510275)
  • Received:2010-01-20 Revised:1900-01-01 Online:2011-01-25 Published:2011-01-25

摘要: 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Epinephelus coioidesTGF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。Realtime 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。

关键词: TGFβ1, 斜带石斑鱼Epinephelus coioides, 克隆, 表达分析

Abstract: 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.

Key words: TGFβ1, Epinephelus coioides, cloning, expression

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