应用Red重组工程技术建立asd基因缺失的大肠杆菌DH10B菌株  

Construction of E.coli DH10B Δasd Using Red Recombineering

作　　者：崔文禹[1] 李山虎[1] 姜飞[1] 吴洋[1] 项勇[2] 周建光[1] 

机构地区：[1]军事医学科学院生物工程研究所,北京100850 [2]沈阳药科大学,辽宁沈阳110016

出　　处：《生物技术通讯》2006年第4期493-495,共3页Letters in Biotechnology

摘　　要：目的:建立一株新遗传表型的大肠杆菌DH10BΔasd菌株。方法:应用pKD46介导的重组系统、kan/kil选择反选择系统、双链线性DNA重组技术和重叠引物介导的DNA重组技术,在菌株DH10B体内,对其染色体上的asd基因进行了基因敲除。结果:建立了一株二氨基庚二酸(DAP)营养缺陷型重组大肠杆菌DH10BΔasd。结论:为进一步建立以大肠杆菌DH10B为载体的DNA疫苗或基因治疗载体奠定了基础。Objective： To construct a new genetype strain E.coli DH10B △asd. Methodes： asd gene was deleted in E. coli DH10B chromosome using homologous recombination system pKD46, kan/kil selection counterselection system, Red mediated linearized double strand DNA homologous recombination and Red mediated recombineering with overlapping single stranded 1）NA oligonucleotides. Results＇. A DAP auxotrophy recombinant bacterium strain was obtained. Conclusion： The recombinant bacterium strain would be able to lay foundation for applications of E.coli DH10B vector in gene therapy and vaccination.

关 键 词：重组质粒 重组工程 同源重组 

分 类 号：Q78[生物学—分子生物学] Q75