Galka Pierre, Jamez Elisabeth, Joachim Gilles, Soumillion Patrice
Institut des Sciences de la Vie, Université catholique de Louvain, Louvain-la-Neuve, Belgium.
PLoS One. 2017 Apr 13;12(4):e0175146. doi: 10.1371/journal.pone.0175146. eCollection 2017.
Incorporation of synthetic degenerate oligonucleotides into plasmids for building highly diverse genetic libraries requires efficient and quantitative DNA manipulation. We present a fast and seamless method for generating libraries of PCR-synthesized plasmids designed with a degenerate sequence and short overlapping ends. Our method called QuickLib should find many applications in synthetic biology; as an example, we easily prepared genetic libraries of Escherichia coli expressing billions of different backbone cyclic peptides.
将合成的简并寡核苷酸整合到质粒中以构建高度多样化的基因文库需要高效且定量的DNA操作。我们提出了一种快速且无缝的方法来生成由PCR合成的质粒文库，这些质粒设计有简并序列和短重叠末端。我们的方法称为QuickLib，在合成生物学中应有许多应用；例如，我们轻松制备了表达数十亿种不同骨架环肽的大肠杆菌基因文库。
