猪SP-A基因启动子区的克隆与序列分析

摘要

为了探明猪SP-A基因的启动子及其转录调控机制,设计6条特异性引物,采用染色体步移、克隆测序以及序列比对分析,从大白猪基因组中扩增出一段长度为1 033 bp的猪SP-A基因的上游序列(DQ985806),其GC含量约为55%.经过生物信息学分析,确定了其转录起始位点及活性区域;发现在转录起始位点上游-33 bp处存在1个TATA-box,另外还发现了GC-box、CAAT-box、GT-box以及转录起始子;该序列还包含Spl、TF和NF-κB等转录因子结合位点以及1个特殊重复序列5'-CATCACTGT-3'.上述试验结果表明,所克隆的1 033 bp的DNA序列是大白猪SP-A基因的启动子区序列.%In order to explore the mechanism of porcine SP-A gene transcription regulation by studying the promoter region of porcine SP-A gene, the upstream sequence of Large White por-cine SP-A gene was attained by Genome Walking PCR,sequencing and sequences alignment anal-ysis. The DNA sequence of 1 033 bp (DQ985806) was amplified. The ratio of G and C in the se-quence was about 55%. The sequence analysis showed that there were a TATA-box at-33 bp and GC-box, CAAT-box, GT-box and transcriptional initiator. Meanwhile, some transcriptional factor binding sites including Spl,TF,NF-κB,and so on were detected. In addition,a special re-petitive sequence of 5'-CATCACTGT-3' was also detected. The results showed that the cloned sequence (1 033 bp) may be considered as the promoter region of Large White porcine SP-A gene.