BACKGROUND Recombinant DNA technology is today a fundamental tool for virtually all biological research fields. Among the many techniques available for the construction of a "custom DNA" molecule, the isothermal in vitro assembly, or Gibson assembly, allows for an efficient, one-step, scarless recombination-based assembly. RESULTS Here, we apply and characterize the use of Gibson assembly for the deletion of DNA sequences around a DNA cut. This method, that we named "Gibson Deletion", can be used to easily substitute or delete one or more restriction sites within a DNA molecule. We show that Gibson Deletion is a viable method to delete up to 100 nucleotides from the DNA ends of a cleavage site. In addition, we found that Gibson Deletion can be performed using single strand DNA with the same efficiency as using double strand DNA molecules. CONCLUSIONS Gibson Deletion is a novel, easy and convenient application of isothermal in vitro assembly, that performs with high efficiency and can be implemented for a broad range of applications.

中文翻译：

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吉布森缺失：等温体外重组的新应用。

背景技术重组DNA技术是当今几乎所有生物学研究领域的基本工具。在许多可用于构建“定制 DNA”分子的技术中，体外等温组装或 Gibson 组装允许高效、一步、无疤痕的重组组装。结果 在这里，我们应用并描述了使用 Gibson 组装来删除 DNA 切割周围的 DNA 序列。这种我们命名为“Gibson 删除”的方法可用于轻松替换或删除 DNA 分子内的一个或多个限制性位点。我们表明，Gibson 删除是一种可行的方法，可以从切割位点的 DNA 末端删除多达 100 个核苷酸。此外，我们发现使用单链 DNA 可以进行吉布森缺失，其效率与使用双链 DNA 分子相同。结论 Gibson 删除是等温体外组装的一种新颖、简单和方便的应用，它执行效率高，可以用于广泛的应用。