乙型肝炎病毒蛋白对核酶剪切作用的影响

Protein action of ribozyme-mediated cleavage on hepatitis B virus in vitro

目的 探讨乙型肝炎病毒蛋白对核酶剪切作用的影响 .方法 利用计算机辅助设计针对乙型肝炎病毒 C基因的三个单一核酶构建 Rz1核酶自剪切转录载体 (p GEMRz1) ,观察单一核酶 (Rz1)对靶 RNA的切割作用及乙型肝炎病毒蛋白对核酶剪切作用的影响 .结果 构建的核酶自剪切转录载体体外转录后 ,在顺式核酶发生自剪切后可将目的核酶正确地释放出来 ,计算机设计的单一核酶体外可剪切靶 RNA;乙型肝炎病毒蛋白对核酶剪切的无明显的抑制或增强作用 ,进一步的结果未观察到乙型肝炎病毒蛋白和靶 RNA的相互作用 .结论 体外转录的核酶可剪切乙肝病毒靶 RNA,在本研究条件下未观察到乙肝病毒蛋白对核酶切割作用的影响 ,从而确保核酶在细胞内能有效发挥作用 .

AIM To detect HBV protein influence on the cleavage activity of ribozyme. METHODS Three single ribozymes against hepatitis B virus core gene were designed with computer, then transctiptional vector of three singleribozymes were constructed, HBV protein influence on the cleavage activity of single ribozyme on target RNA observed, and HBV protein interaction with target RNA further analysed. RESULTS Single antiviral ribozyme(Rz1) could cleave its target RNA efficiently, HBV proteins had no effects on ribozyme mediated cleavage on HBV. Further results indicated that HBV proteins did not interact with target RNA in our experiment. CONCLUSION HBV proteins has no effects on ribozyme mediated cleavage on HBV.