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为探索有效的登革热治疗方法 ,根据DEN - 2基因结构的特点 ,按照锤头状核酶的结构模型和作用模式 ,设计、合成了针对 172～ 174GUC位点的核酶基因 ;其末端分别加上SacI和SalI接头 ,定向插入质粒pGEM - 3zf (+ )的SacI和SalI位点之间 ;其引导序列中的PvuII位点 ,提供了快速简便的核酶克隆的鉴定方法。PvuII酶切鉴定的核酶基因克隆经序列分析 ,结果与预期完全一致。证明得到了特异切割DEN - 2基因的核酶基因克隆 ,为进一步核酶抗DEN - 2的研究打下了基础
In order to explore an effective dengue treatment method, according to the structure of DEN - 2 gene, the ribozyme gene targeting 172-174GUC site was designed and synthesized in accordance with the structural model and mode of action of hammerhead ribozyme. The SacI and SalI adapters were inserted between the SacI and SalI sites of the plasmid pGEM - 3zf (+). The guide sequence of the PvuII site provided a rapid and convenient method for the identification of ribozyme clones. The ribozyme gene clones identified by PvuII digestion were sequenced and the results were exactly the same as expected. It was proved that the ribozyme gene cloned by specific cleavage of DEN - 2 gene laid the foundation for the further research of ribozyme against DEN - 2.