Modular Cloning by Golden Gate Assembly and Possible Application in Pathway Design

• DOI: 10.2533/chimia.2023.437
• Publication Date: 2023-06-28T03:36:58Z
• Authors: Zrinka Raguz Nakic; Christin Peters
• Keywords: Cloning (programming); Synthetic Biology; Golden gate

Preparation of expression vectors using conventional cloning strategies is laborious and not suitable for the design metabolic pathways or enzyme cascades, which usually requires preparation a vector library to identify productive clones. Recently, Modular Cloning as novel technique in synthetic biology has been developed. relies on Golden Gate assembly supports individual one-step one-pot reactions, thus allowing rapid generation libraries. A number toolkits specific applications established, providing collection distinct genetic elements such promoters, ribosome binding sites tags, that can be combined individually defined fusion sites. successfully applied generate various strains producing value-added compounds. This was achieved by orchestrating complex involving up 20 enzymes. Due novelty approach, industrial are still rare. In addition, some limited due lack high-throughput screening methods. shifts bottleneck from capacity needs addressed future developments pave path establishment applications.