Javorova Rachel, Rezuchova Bronislava, Feckova Lubomira, Novakova Renata, Csolleiova Dominika, Kopacova Maria, Patoprsty Vladimir, Opaterny Filip, Sevcikova Beatrica, Kormanec Jan
Institute of Molecular Biology, Slovak Academy of Sciences, Bratislava 845 51, Slovak Republic.
Institute of Chemistry, Slovak Academy of Sciences, Bratislava 845 38, Slovak Republic.
J Biotechnol. 2024 Sep 10;392:128-138. doi: 10.1016/j.jbiotec.2024.07.007. Epub 2024 Jul 14.
We have created a novel synthetic biology expression system allowing easy refactoring of biosynthetic gene clusters (BGCs) as monocistronic transcriptional units. The system is based on a set of plasmids containing a strong kasOp* promoter, RBS and terminators. It allows the cloning of biosynthetic genes into transcriptional units kasOp*-gene(s)-terminator flanked by several rare restriction cloning sites that can be sequentially combined into the artificial BGC in three compatible Streptomyces integration vectors. They allow a simultaneous integration of these BGCs at three different attB sites in the Streptomyces chromosome. The system was validated with biosynthetic genes from two known BGCs for aromatic polyketides landomycin and mithramycin.
我们创建了一种新型合成生物学表达系统，可轻松将生物合成基因簇（BGC）重构为单顺反子转录单元。该系统基于一组含有强kasOp启动子、核糖体结合位点（RBS）和终止子的质粒。它允许将生物合成基因克隆到转录单元kasOp-基因（多个）-终止子中，两侧有几个稀有限制性克隆位点，这些位点可在三种兼容的链霉菌整合载体中依次组合成人工BGC。它们允许这些BGC同时整合到链霉菌染色体的三个不同attB位点。该系统已用来自两种已知芳香族聚酮化合物——兰卡霉素和光神霉素的生物合成基因进行了验证。
