›› 2011, Vol. 38 ›› Issue (3): 87-91.

• 生物技术 • 上一篇    下一篇

绵羊甘露聚糖结合凝集素基因启动子区的克隆与序列分析

余鹏1，王遵宝2，吴洪宾2，班谦2，赵宗胜1,2   

  1. （1.石河子大学生命科学学院，新疆石河子 832003； 2.石河子大学动物科技学院，新疆石河子 832003）
  • 收稿日期:1900-01-01 修回日期:1900-01-01 出版日期:2011-03-20 发布日期:2011-03-20
  • 通讯作者: 赵宗胜

Cloning and Sequence Analysis of the Promoter Region of Sheep Mannan-binding Lectin Gene

YU Peng1，WANG Zun-bao2，WU Hong-bin2，Ban qian2，ZHAO Zong-sheng1,2   

  1. （1.College of Life Science, Shihezi University, Shihezi 832003, China；2.College of Animal Science and Technology, Shihezi University, Shihezi 832003, China）
  • Received:1900-01-01 Revised:1900-01-01 Online:2011-03-20 Published:2011-03-20
  • Contact: ZHAO Zong-sheng

摘要： 为了探讨绵羊甘露聚糖结合凝集素（mannan-binding lectin，MBL）基因启动子区活性及转录调控机制，本试验根据已提交的绵羊MBL序列设计3条特异性引物，采用热不对称交错PCR(thermal asymmetric interlaced PCR，TAIL-PCR)技术扩增获得绵羊MBL基因的启动子区序列。经生物信息学分析，确定了其转录活性区域，发现绵羊MBL基因启动子无TATA框，但其存在多个PEA3和Spi-1/PU.1转录因子结合位点，二者同属Ets家族,且参与绵羊MBL基因转录起始复合体的形成。此外，该序列还包含Sp1、GATA-1、TCF/LEF等其他转录因子结合位点。结果表明，试验成功克隆获得了绵羊MBL基因的启动子区序列，为后期MBL基因启动子区活性及其表达调控机制和甲基化研究奠定基础

关键词: 绵羊; MBL; 启动子区; 克隆; TAIL-PCR; 序列分析

Abstract: In order to investigate the promoter region activity and transcriptional control mechanism of ovine mannan-binding lectin (MBL) gene, the thermal asymmetric interlaced PCR (TAIL-PCR) technology was adopted in this study. Meanwhile, three specific primers were designed according to the MBL sequence submitted on sheep, and we ultimately got the promoter sequences of the sheep MBL gene.By the bioinformatics analysis, we defined its transcriptional activity region, and found that the promoter of sheep MBL gene didn’t have TATA-box, however, there were several binding sites between PEA3 and Spi-1/PU1 transcription factor, which both belonged to the Ets family and were involved in the formation of transcription initiation complex formation in ovine MBL gene. In addition, the sequence also contained Sp1, GATA-1, TCF/LEF and other binding sites of transcription factors. The clone of promoter region sequence of ovine MBL gene in this study, made a foundation for the latter studies on the promoter region activity and its expression and control mechanisms, even on the methylation of the sheep MBL gene.

Key words: sheep; MBL; promoter region; clone; TAIL-PCR; sequence analysis

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