The construction of large DNA molecules that encode pathways, biological machinery, and entire genomes has been limited to the reproduction of natural sequences. However, now that robust methods for assembling hundreds of DNA fragments into constructs >20 kb are readily available, optimization of large genetic elements for metabolic engineering purposes is becoming more routine. Here, various DNA assembly methodologies are reviewed and some of their potential applications are discussed. We tested the potential of DNA assembly to install rational changes in complex biosynthetic pathways, their potential for generating complex libraries, and consider how various strategies are applicable to metabolic engineering.

编码途径，生物机制和整个基因组的大型DNA分子的构建仅限于天然序列的复制。但是，现在已经可以使用健壮的方法将数百个DNA片段组装成大于20 kb的构建体，为代谢工程目的而优化大型遗传元件变得越来越常规。在这里，审查了各种DNA组装方法，并讨论了它们的一些潜在应用。我们测试了DNA组装在复杂生物合成途径中安装合理变化的潜力，它们在生成复杂文库中的潜力，并考虑了各种策略如何适用于代谢工程。