Wang Yi, Zhang Zhong-Tian, Seo Seung-Oh, Choi Kijoong, Lu Ting, Jin Yong-Su, Blaschek Hans P
Department of Food Science and Human Nutrition, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA; Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
Department of Chemistry, University of Illinois at Urbana-Champaign, Urbana, IL 61801, USA.
J Biotechnol. 2015 Apr 20;200:1-5. doi: 10.1016/j.jbiotec.2015.02.005. Epub 2015 Feb 11.
The anaerobic spore-forming, gram-positive, solventogenic clostridia are notorious for being difficult to genetically engineer. Based on CRISPR/Cas9 assisted homologous recombination, we demonstrated that clean markerless gene deletion from the chromosome can be easily achieved with a high efficiency through a single-step transformation in Clostridium beijerinckii NCIMB 8052, one of the most prominent strains for acetone, butanol and ethanol (ABE) production. This highly efficient genome engineering system can be further explored for multiplex genome engineering purposes. The protocols and principles developed in this study provided valuable references for genome engineering in other microorganisms lacking developed genetic engineering tools.
厌氧产芽孢、革兰氏阳性、产溶剂梭菌因难以进行基因工程改造而声名狼藉。基于CRISPR/Cas9辅助的同源重组，我们证明，在拜氏梭菌NCIMB 8052（丙酮、丁醇和乙醇（ABE）生产中最突出的菌株之一）中，通过单步转化可以轻松高效地从染色体上实现无标记基因的无痕删除。这种高效的基因组工程系统可进一步用于多重基因组工程目的。本研究中开发的方案和原理为缺乏成熟基因工程工具的其他微生物的基因组工程提供了有价值的参考。
