Recombineering:: In vivo genetic engineering in E-coli, S-enterica, and beyond

被引:163
作者
Sawitzke, James A. [1 ]
Thomason, Lynn C. [1 ]
Costantino, Nina [1 ]
Bubunenko, Mikhail [1 ]
Datta, Simanti [1 ]
Court, Donald L. [1 ]
机构
[1] NCI, Frederick, MD 21701 USA
来源
ADVANCED BACTERIAL GENETICS: USE OF TRANSPOSONS AND PHAGE FOR GENOMIC ENGIEERING | 2007年 / 421卷
关键词
D O I
10.1016/S0076-6879(06)21015-2
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
Recombineering, in vivo genetic engineering with short DNA homologies, is changing how constructs are made. The methods are simple, precise, efficient, rapid, and inexpensive. Complicated genetic constructs that can be difficult or even impossible to make with in vitro genetic engineering can be created in days with recombineering. DNA molecules that are too large to manipulate with classical techniques are amenable to recombineering. This technology utilizes the phage lambda homologous recombination functions, proteins that can efficiently catalyze recombination between short homologies. Recombineering can be accomplished with linear PCR products or even single-stranded oligos. In this chapter we discuss methods of and ways to use recombineering.
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页码:171 / 199
页数:29
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