摘要
目的观察丁型肝炎病毒2种基因组核酶(HDVgenomicribozyme,g.Rz)是否存在反式剪切活性。方法合成g.Rz74和g.Rz55的cDNA ,克隆入质粒pGEM 4Z ,体外转录获取核酶RNA ;同时从质粒pRz277B上转录出同源性底物RNA ,以α 32p UTP标志 ;再将核酶与底物按比例混合 ,在一定条件下观察是否出现剪切作用。结果启动反应30min后 ,可以观察到剪切产物 ,90min后剪切反应更加明显。结论 g.Rz74和g.Rz55均具有反式剪切活性。
Objective To study the trans cleaving activity of 2 genomic ribozymes (g.Rz) of hepatitis delta virus (HDV). Methods The cDNA of g.Rz 74 and g.Rz 55 was synthesized and cloned into plasmid pGEM 4Z and g.Rz RNA was obtained through transcription in vitro. Meanwhile, homogenous substrate RNA was transcribed from plasmid pRz277B and labelled with α 32p UTP. The ribozymes were mixed with the substrate in appropriate proportion under certain conditions to observe whether there existed any trans cleaving activity. Results The products of trans cleavage were obtained 30 minutes after the initiation of the reaction and the reaction was further intensified 90 minutes later. Conclusion Both g.Rz 74 AND g.Rz 55 possess trans cleaving activity.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2000年第4期305-306,共2页
Journal of Third Military Medical University
基金
国家自然科学基金!资助项目(39600031)
关键词
丁型肝炎病毒
反式剪切
基因组核酶
HDV ribozyme
trans cleavage
genome
transcription in vitro
