摘要
Ribozyme是一类具有生物催化功能的RNA,它能定点切割RNA靶分子,从而达到有效地阻断基因表达的目的。本文依“锤头结构”原理设计合成了一个38个核苷酸的Ribozyme,能在体外定点切割甲型肝炎病毒的RNA片段(链长为123核苷酸残基,切削点对应于甲肝病毒RNA的559位)。测定了切割反应与温度、Ribozyme浓度和底物浓度的关系,其Km和kcat分别为0.63μmol/L和0.12min^(-1)。Mg^(2+)或Ca^(2+)均可作为参与该催化反应所必需的金属离子。
A 123mer RNA fragment containing 533-633 sequence of hepatitis A virus ( HAV ) and used as substrate was obtained by transcription from DNA plasmid constructed with pHAVL1307 and pTZl9R ( Pharmacia ). A 38mer of oligoribonucleotlde used as nbozyme was designed according to the 'hammerhead structure' suggested by Symons' group and synthe sized chemically. The 123mer substrate derived from HAV can be clea-ved site-specit'ically by the 38mer ribozyme in uitro, The relationship between cleavage efficiency and temperature, ribozyme and substrate concentrations were studied. The Km and kcat values for the reaction are 0.63βmol/L and 0.12min-1 respectively. Either Mg2+ or Ca2+ is nece-ssary in the catalytic reaction.
出处
《病毒学报》
CAS
CSCD
北大核心
1993年第1期1-6,共6页
Chinese Journal of Virology
基金
国家863高科技生物技术领域资助
