FUS1基因慢病毒载体的构建  

Construction of lentivirus vector containing FUS1 gene

作　　者：张宝[1] 霍霞[1] 彭琳[1] 齐宗利[1] 徐锡金[1] 李燕[1] 郑良楷[1] 丘波[1] 

机构地区：[1]汕头大学医学院中心实验室

出　　处：《中华肿瘤防治杂志》2008年第4期241-243,共3页Chinese Journal of Cancer Prevention and Treatment

基　　金：中国博士后科学基金(2005038187);国家自然科学基金(30600733)

摘　　要：目的:构建FUS1基因慢病毒载体,包装病毒,体外高效感染细胞。方法:采用PCR技术从人脐带来源的间充质干细胞中扩增出FUS1基因,并将其接入慢病毒载体pSL6-IRES-EGFP中,经PCR、酶切和测序鉴定后,与包装质粒共转染293T细胞,制备慢病毒。将病毒转染BEL7404和DLD-1细胞,观察病毒转染效果。结果:经过PCR测序鉴定,克隆了pSL6-FUS1-IRES-EGFP重组子;所包装的病毒对细胞的感染效率>90%。结论:成功地克隆FUS1基因于慢病毒载体,制备了高效感染细胞的慢病毒。OBJECTIVE： To construct the lentivirus vector containing FUS1 gene and prepare lentivirus for high transfection. METHODS： The full length of FUS1 gene was amplified by the method of PCR technique from the total RNA of the umbilical mesenchymal stem cells. FUS1 gene was cloned into pSL6-IRES-EGFP vector and identified with PCR, digestion and sequencing. The recombinant plasmid of pSL6-FUSI-IRES-EGFP was packaged in 293T cells and the lentivirus was collected. The infection effect was observed by transfection into BEL7404 and DLD-1 cells. RESULTS： The plasmid of pSL6- FUSI-IRES-EGFP was obtained and identified by means of PCR, digestion, sequencing. The lentivirus can infect cells more than 90%. CONCLUSION： The recombinant lentivirus vector of FUS1 was successfully cloned and the preparation of lentivirus may high effectively transfect cells.

关 键 词：基因 肿瘤抑制 基因 FUS1 慢病毒属 转染 

分 类 号：R346[医药卫生—基础医学]