摘要
目的:构建并表达衣壳蛋白表面展示有精氨酸-甘氨酸-天冬氨酸(RGD)序列的MS2噬菌体样颗粒,以期获得肿瘤细胞靶向性RNA递送载体。方法:用分子克隆技术将RGD4C编码序列插入MS2衣壳蛋白编码基因,然后构建到p ETDuet-1质粒多克隆位点MCS1上;将含有MS2噬菌体包装位点的pre-let-7编码基因构建到p ETDuet-1质粒多克隆位点MCS2上;重组质粒转化大肠杆菌,表达产物纯化后经琼脂糖凝胶电泳、电子显微镜鉴定。结果和结论:采用分子克隆技术和原核细胞单质粒双表达系统获得表面展示有RGD序列,内部包装有pre-let-7 RNA的MS2噬菌体样颗粒,为体内和体外研究其肿瘤细胞靶向递送和功能奠定了基础。
Objective: To construct and express MS2 bacteriophage like particles containing RGD sequence as adelivery vector for tumor target therapy.Methods: The MS2 capsid encoding sequence containing RGD4 C se-quence was cloned to MCS1 of plasmid p ETDuet-1,and the pre-let-7 encoding sequence was cloned to MCS2 ofplasmid p ETDuet-1.The recombinant plasmids were transformed into E.coli.The purified bacteriophage like parti-cles were analyzed by 1% agarose gel electrophoresis with ethidium bromide staining and by transmission electronmicroscopy.Results & Conclusion: We successfully produced RGD modified MS2 bacteriophage like particlespackaging pre-let-7 RNA in E.coli,which provides basis for further study in vitro and in vivo.
引文
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