Gateway Recombinational Cloning.
Gateway Recombinational Cloning.
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DOI：
10.1101/pdb.top094912
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发表时间：
2018-01-02
影响因子：
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通讯作者：
Walhout AJM
中科院分区：
文献类型：
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作者：
Reece-Hoyes JS;Walhout AJM
Gateway 重组克隆系统旨在使用相同的酶以标准化方式并行克隆多个 DNA 片段（例如，以 96 孔格式）。 Gateway 克隆基于噬菌体 λ 进出大肠杆菌基因组的高度特异性整合和切除反应。由于重组位点（“att”位点）比限制性位点长得多（25-242 bp），因此它们极不可能偶然出现在 DNA 片段中。因此，相同的重组酶可用于在平行反应中稳健地克隆许多不同大小的不同片段。
The Gateway recombinatorial cloning system was developed for cloning multiple DNA fragments in parallel (e.g., in 96-well formats) in a standardized manner using the same enzymes. Gateway cloning is based on the highly specific integration and excision reactions of bacteriophage λ into and out of the Escherichia coli genome. Because the sites of recombination (“att” sites) are much longer (25–242 bp) than restriction sites, they are extremely unlikely to occur by chance in DNA fragments. Therefore, the same recombination enzyme can be used to robustly clone many different fragments of variable size in parallel reactions.