表面展示RGD模体重组噬菌体的构建和鉴定  

Construction and identification of a recombinant phage vector display RGD motif for gene delivery

作　　者：夏文龙[1] 余树培 郭长明[2] 朱善元[2] 张鑫宇[1] 夏晓莉[1] 孙怀昌[1] 

机构地区：[1]扬州大学兽医学院/江苏省重要动物疫病与人兽共患病协同创新中心,江苏扬州225009 [2]江苏农牧科技职业学院/江苏省兽用生物制药高技术研究重点实验室,江苏泰州225300

出　　处：《扬州大学学报（农业与生命科学版）》2017年第4期9-13,24,共6页Journal of Yangzhou University：Agricultural and Life Science Edition

基　　金：江苏省自然科学基金资助项目(BK20151576);江苏高校优势学科建设工程项目(PAPD);江苏省农业科技自主创新资金项目[CX(14)2087];江苏农牧科技职业学院科研项目(NSFPT201631)

摘　　要：将RGD多肽编码序列克隆至辅助噬菌体M13KO7 PⅢ基因中,获得重组载体M13KO7-RGD,包装噬菌粒pAAV-GFP后获得重组噬菌体颗粒,通过透射电子显微镜观察形态,Western-blot鉴定PⅢ-RGD融合序列的表达。将其转导不同哺乳动物细胞,荧光显微镜观察报告基因的表达。结果表明:成功获得展示RGD模体的重组噬菌体颗粒,且具有典型丝状噬菌体形态特征,转导不同细胞系后有绿色荧光蛋白表达,提示该重组噬菌体能够导入哺乳动物细胞,为动物基因工程疫苗载体的研发提供新的思路。The coding sequence of RGD peptide was cloned to P Ⅲ gene of helper phage M13KO7,obtaining recombinant vector M13KO7-RGD.Recombinant phage particles were prepared after packaging pAAV-GFP phagemid by M13KO7-RGD.Morphologies was observed by transmission electron microscopy,and expression of P Ⅲ-RGD was identified by Western-blot.Phage particles were transduced to different mammalian cells and expression of reporter gene was observed by fluorescence microscope.Results showed that recombinant phage particles displayed RGD motif were obtained successfully,which had typical characteristics of filamentous phage.Green fluorescent protein was expressed after transduction of recombinant phage particles to different animal cells.These results indicated that the recombinant phage can be successfully transformed into mammalian cells,which provides a new idea for research and development of animal genetic engineering vaccine vectors.

关 键 词：重组噬菌体 RGD模体 基因导入 

分 类 号：Q518.1[生物学—生物化学]