MacDonald James T, Siciliano Velia
Centre for Synthetic Biology and Innovation, Imperial College, South Kensington Campus, London, SW7 2AZ, UK.
Department of Medicine, Imperial College, South Kensington Campus, London, SW7 2AZ, UK.
Methods Mol Biol. 2017;1651:249-262. doi: 10.1007/978-1-4939-7223-4_18.
Recently developed DNA assembly methods have enabled the rapid and simultaneous assembly of multiple parts to create complex synthetic gene circuits. A number of groups have proposed the use of computationally designed orthogonal spacer sequences to guide the ordered assembly of parts using overlap-directed or homologous recombination-based methods. This approach is particularly useful for assembling multiple parts with repetitive elements. Orthogonal spacer sequences (sometimes called UNSs-unique nucleotide sequences) also have a number of other potential uses including in the design of synthetic promoters regulated by novel regulatory elements.
最近开发的DNA组装方法能够快速同时组装多个部件，以创建复杂的合成基因电路。许多研究小组提出使用经过计算设计的正交间隔序列，通过基于重叠导向或同源重组的方法来指导部件的有序组装。这种方法对于组装具有重复元件的多个部件特别有用。正交间隔序列（有时称为UNS——独特核苷酸序列）还有许多其他潜在用途，包括设计由新型调控元件调控的合成启动子。
