【摘要】 目的:克隆大鼠睾丸组织uPA基因的启动子区并对该序列进行分析。方法:从大鼠睾丸组织中提取基因组DNA,以其为模板设计uPA基因引物,运用降落PCR法扩增uPA基因5’端上游的真核转录调控序列。测序得到的PCR产物用启动子区的分析软件分析,并与其DNA序列进行比对。结果:得到的uPA基因长度为1572bp(登录号X65651)。经软件分析:该序列包含uPA基因完整的开放阅读框(ORF)、21bp的外显子部分,1551bp区域为转录起始的上游部分在其5’端UTR区的-30bp位置有一个不典型的TATA盒,其上游有非常明显的GC盒及启动子区常见的AP1(activeprotein1)、SP1等结合位点。结论:成功克隆了大鼠睾丸组织uPA基因启动子区。分析表明,该片段包含真核转录调控区域、不典型的TATA盒、典型的GC盒及启动子区所常见的AP1、SP1等结合区域。更多还原

【Abstract】 Objective:To clone and analyze the promoter sequence of rat urokinase plasminogen activator(uPA)protein gene.Methods:The genomic DNA was extracted from rat testicle tissue.According to uPA,the gene upper and lower primer of uPA gene were designed and synthesized,then touch-down PCR were performed.After proper purification,the PCR product was sequence,analyzed with the promoter prediction software and compared with the DNA sequence of rat uPA.Results:The cloned uPA gene was about 1 572 bp in length,Which contained a full open-reading frame with 21 bp in length exons,and the upper region of transcriptional start is1 551 bp in length which is eucaryou transcriptional control area.The 5’UTR has a promoter region including a non-conspicu-ous TATA-box.Not only the GC-box binding region was found in this gene,but also active protein 1(AP1)and SP1 were seen in other regions.Conclusion:A 1 572 bp uPA gene fragment(GenBank accession NO.X65651)was obtained from rat genomic DNA library,containing eucaryou transcriptional control area with a promoter region,non-conspicuous TATA-box,GC-box and 1 extrons.A TATA-box was located at the upper-30 region.更多还原