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DNA cloning using in vitro site-specific recombination

Hartley, J L ; Temple, G F ; Brasch, M A

Genome research, 2000-11, Vol.10 (11), p.1788-1795 [Periódico revisado por pares]

United States: Cold Spring Harbor Laboratory Press

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Título:
DNA cloning using in vitro site-specific recombination
Autor: Hartley, J L ; Temple, G F ; Brasch, M A
Assuntos: Animals ; Artificial Gene Fusion ; Cloning, Molecular - methods ; DNA - genetics ; DNA, Bacterial - genetics ; Escherichia coli - genetics ; Gene Expression ; Genetic Vectors - chemical synthesis ; Genetic Vectors - metabolism ; Humans ; In Vitro Techniques ; Methods ; Polymerase Chain Reaction ; Protein Biosynthesis ; Proteins - genetics ; Recombination, Genetic
É parte de: Genome research, 2000-11, Vol.10 (11), p.1788-1795
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content type line 23
Corresponding author.
Descrição: As a result of numerous genome sequencing projects, large numbers of candidate open reading frames are being identified, many of which have no known function. Analysis of these genes typically involves the transfer of DNA segments into a variety of vector backgrounds for protein expression and functional analysis. We describe a method called recombinational cloning that uses in vitro site-specific recombination to accomplish the directional cloning of PCR products and the subsequent automatic subcloning of the DNA segment into new vector backbones at high efficiency. Numerous DNA segments can be transferred in parallel into many different vector backgrounds, providing an approach to high-throughput, in-depth functional analysis of genes and rapid optimization of protein expression. The resulting subclones maintain orientation and reading frame register, allowing amino- and carboxy-terminal translation fusions to be generated. In this paper, we outline the concepts of this approach and provide several examples that highlight some of its potential.
Editor: United States: Cold Spring Harbor Laboratory Press
Idioma: Inglês

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DNA cloning using in vitro site-specific recombination

Hartley, J L ; Temple, G F ; Brasch, M A

United States: Cold Spring Harbor Laboratory Press

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