Projects per year
cloning) has been described by several research groups, but the optimal design of
cohesive DNA ends for multigene assembly remains elusive. Here, we use two
model constructs based on expression of gfp and a four-gene pathway that
produces β-carotene to optimize assembly junctions and the uracil excision
protocol. By combining uracil excision cloning with a genomic integration
technology, we demonstrate that up to six DNA fragments can be assembled in a
one-tube reaction for direct genome integration with high accuracy, greatly
facilitating the advanced engineering of robust cell factories.
- Molecular cloning
- DNA assembly
- Uracil excision cloning
- Genome engineering
BacTory: Biorefinery Training Platform: Bacterial Factories for Sustainable Chemical and Drug Production
Molin, S., Nørholm, M., Cavaleiro, M., Machado, H., Wendel, S., van der Helm, E., Bojanovic, K., Vazquez Albacete, D., D'Arrigo, I., Li, S., Yang, X., Redl, S. M. A., Pogrebnyakov, I., Lehning, C. E., Giubergia, S., Calero Valdayo, P. M. & Lohmann, R.
01/10/2012 → 31/01/2017