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目的探讨多聚嘧啶序列结合蛋白(polypyrimidine tract binding protein,PTB)对乙型肝炎病毒转录后调节元件(HBV posttranscriptional regulatory element,HPRE)功能的影响。方法采用pDM138-HPRE CAT(chloramphenicol acetyltransferase)报告体系,通过质粒瞬时转染细胞的方式来探讨PTB对HPRE功能的影响。结果CAT活性检测表明:PTB的过表达使HPRE介导的CAT表达活性下降,并且呈剂量依赖性。当去除质粒上游剪接供体位点时,PTB仍然可以降低CAT的活性。结论PTB抑制HPRE的功能,并且这种抑制作用不依赖于上游剪接供体位点。
Objective To investigate the effect of polypyrimidine tract binding protein (PTB) on the function of Hepatitis B virus posttranscriptional regulatory element (HPRE). Methods The pDM138-HPRE CAT (chloramphenicol acetyltransferase) reporter system was used to investigate the effect of PTB on the function of HPRE by transient transfection of plasmids into cells. Results CAT activity assay showed that PTB overexpression reduced the activity of HPRE-mediated CAT expression in a dose-dependent manner. PTB can still reduce CAT activity when removing donor splice donor sites upstream of the plasmid. Conclusion PTB inhibits the function of HPRE, and this inhibitory effect does not depend on the upstream splice donor site.