摘要
目的:扩增乙酰肝素酶(heparanase,HPSE)基因启动子核心片段,并进行序列分析。方法:提取人类基因组DNA,PCR扩增HPSE基因启动子,T-A克隆、酶切鉴定,然后进行测序比对,分析其中转录因子结合位点。结果:从人类基因组DNA中扩增出HPSE基因启动子片段,酶切鉴定与预期结果吻合,长度为561bp,该启动子碱基序列与数据库GenBank完全一致,并含有3个Sp1、4个ERE和2个ERG1转录因子结合位点。结论:成功克隆了HPSE核心启动子,并含有维持HPSE转录活性的转录因子结合位点,为后续研究奠定了基础。
Objective ： To amplify the human heparanase （ HPSE ） gene core promoter for sequence analysis. Methods ： By extraction of genomie DNA,the human HPSE gene promoter was amplified and identified by T- A cloning, restriction endonuelease and sequencing for determining the transcription factor binding sites in the amplified sequence. Results： A segment of human HPSE gene promoter was amplified from the genomic DNA and its size（561bp in length）was identical to estimated results. The DNA sequence of amplified HPSE gene promoter was accordant with that of GenBank data,including three Spl, four Ets-relevant elements （ ERE ） and two early growth response gene 1 （ EGR1 ） binding sites. Conclusion ： The human HPSE gene core promoter, containing the binding sites of some major transcription factor that tuaintains the activity of transcription,was successfully cloned,which will lay the foundation tor further study in this field.
出处
《皖南医学院学报》
CAS
2009年第5期319-322,共4页
Journal of Wannan Medical College
基金
安徽省教育厅自然科学基金项目(2003kj312)
皖南医学院附属弋矶山医院博士基金项目(2003B001)
