抗HPV11 E2核酶的计算机设计

 借助计算机软件分析 ,设计出能特异性切割HPV11型 6 4 4ntE2mRNA的核酶 (ribozyme) .遵循Symon′s锤头状核酶结构和GUX剪切位点原则 ,靶序列存在 32个这样的剪切位点 .通过计算机软件分析出核酶的最佳剪切位点 ,并对底物及核酶的二级结构进行预测及进行相应基因生物学功能和基因同源性分析 ,筛选出 2个锤头结构核酶 .针对这两位点设计的核酶分别命名为RZ2 777和RZ32 81.计算机分析显示 ,两核酶与底物切点两翼碱基形成锤头状结构 ,切点所在基因序列具有相对松弛的二级结构 ,位于该基因重要生物功能区内 ,是核酶的理想攻击区域 .通过基因库检索 ,在已知人类基因排除了与上述两核酶切点两翼碱基有基因同源性序列的可能性 .将两核酶用于体外剪切实验取得了良好的实验结果 ,认为借助计算机分析可帮助尽快从多个剪切位点选择出最适核酶

Computer Design of Anti-HPV11 E2 Ribozyme

HPV11 E2 mRNA was taken as the target RNA.Ribozyme were designed accounting to the hammerhead structure described by Symons.Computer was used to analyze the possible secondary cleavage sites on HPV11 E2 mRNA and to predict the secondary structure of substrate and ribozymes.According to the theory of Symon's hammerhead ribozyme,there are 32 targetting sites sequences.The secondary structure of HPV11 mRNA from nt2510 to nt3518 was relatively stable.The region had important biological functions and was the ideal attacking region for ribozyme.Two ribozymes targeting nt2777 and nt3281 on the HPV11 mRNA were designed and were named RZ2777 and RZ3281 respectively.No analogous sequence susceptible to the designed ribozyme was found in the mRNA of others genes in known human GenBank through computer search.Not all GUX or CUX could be taken as the cleavage site of ribozyme.The nt2777 and nt3281 on HPV11 mRNA are the most ideal attacking sites for ribozymes.So computer analysis can assist to select the optimum ribozyme design from a large number of possible cleavage sites.