抗HPV11 E2核酶的计算机设计

Computer design of anti-HPV11 E2 ribozyme

借助计算机软件分析 ,设计出能特异性切割HPV11型 644ntE2mRNA的核酶。遵循Symons锤头状核酶结构和GUX剪切位点原则 ,靶序列存在 3 2个剪切位点 ,通过计算机软件分析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RZ3 2 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 ,在已知人类基因中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。并非所有的GUX位点 (X :C、U、A)或CUX均可作为核酶的最佳剪切位点 ,计算机分析可帮助尽快选择出最适核酶 ,指导进行核酶体外转录载体构建并进行切割反应 ,为下一步将核酶用于细胞内和体内试验打下基础。

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 structures of substrate and ribozymes. According to the theory of Symons headhammer ribozyme, there are 32 sites to targetting sequences. The secondary structures of HPV11 mRNA from nt2510 to nt3518 were relatively stable. The region was of important biological function 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 of substrate which combined with ribozyme was found in the mRNA of others genes in known Human Gene Bank through computer probe. Not all GUX or CUX could be taken as the cleavage site of ribozyme. The nt2777 and nt3281 onr HPV11 mRNA are the most ideal attacking sites for ribozymes. So computer analysis was used to select the optium ribozyme as soon as possible and to lay a solid foundation for using ribozyme in vivo.