The aims of this study were to clone and analyze the promoter sequence of porcine SelS gene,and preliminary investigate the effect of transcription factors binding sites on the expression of porcine SelS gene. The promoter sequence of porcine SelS gene was cloned using SON-PCR,the characterization of this promoter was predicted using online tools such as PromoterScan and Promoter 2. 0. The effect of NF-kappaB on the promoter activity was detected in PK15 cells after LPS stimulation. An approximate 3 kb promoter sequence was acquired and the homology of partial promoter sequence was only 7％-51％ among four species including pig, human, cattle and mouse. The transcription start site of porcine SelS gene located at -398 bp, the conservative transcription factors binding sites between pig and human SelS gene included NF-kappaB,CCAAT box,SPI and USF,except the typical TATA box. Cell experiments showed NF-kappaB could up-regulate the expression of porcine SelS gene. The results indicate that the sequence homology of SelS gene is low among examined species, but the transcription factors are very conservative between pig and human. The LPS induction experiment suggest that the expression of porcine SelS gene may be regulated by NF-kappaB.%本研究旨在克隆和分析猪硒蛋白S基因(Selenoprotein S,SelS)启动子序列,并初步探讨潜在转录因子结合位点对其表达的影响.通过SON-PCR技术克隆猪SelS基因启动子序列,利用PromoterScan、Promoter 2.0等在线工具预测其启动子特征,利用细菌脂多糖(Lipopolysaccharides,LPS)刺激PK15细胞,研究NF-kappaB转录因子对猪SelS基因启动子活性的影响.试验获得了猪SelS基因约3kd的启动子序列,部分序列比对发现猪、人、牛和小鼠物种间相似性仅7%～51%.预测猪SelS基因转录起始位点在-398 bp,猪和人SelS基因启动子存在系列保守的转录因子结合位点,包括NF-kappaB、CCAAT box、SP1、USF等,但均未发现典型的TATA box.细胞试验表明,NF-kappaB转录因子可以上调猪SelS基因的表达.结果提示,物种间SelS基因启动子相似性较低,但猪和人SelS基因的转录因子非常保守,LPS诱导试验提示,猪SelS基因表达可能受NF-kappaB转录因子的调控.
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