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。
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/PU.1 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.
出处
《中国畜牧兽医》
CAS
北大核心
2011年第3期87-92,共6页
China Animal Husbandry & Veterinary Medicine
基金
国家自然基金项目(30960247)
