 Hello, my name is Jianli. I am a postdoc at Northwestern University. In this video, I would like to introduce you our recent published paper in biotechnology and bioengineering. The title is Establishing a High-Yielding Structurizing Spaced Self-Read Protein Synthesis System. In this work, we aim to develop a self-read system for the expression of high-GC content genes. To this end, we choose the structurizing species as our target students because structurizing students have high-GC content genomes. They are featured by many complex natural products, for example the antibiotic biosynthetic gene clusters. To establish a system for expressing GC-rich genes is important because in the future with this system, people can easily express the high-GC content biosynthetic gene clusters from structurizing. For the natural products synthesis in a self-read environment, this system is highly open and controllable, allowing for easy manipulation, monitoring, sampling, and optimization. Among numerous structurizing strains, we choose the structurizing leave-dance as our initial strain, since this strain has been widely used for enviable heterogeneous protein expression. In addition, we use the code-optimized EGFP as a report protein. In order to make the cell light set, we grow the cells to its exponential phase followed by cell collection, disruption, and clarification. Then, we assembled the cell-free protein synthesis reaction in a tube and optimized various parameters. After optimization, we achieved the EGFP yield of more than 50 micrograms per mil in a batch reaction, which was further increased to nearly 300 micrograms per mil with a semi-continuous reaction after 48 hours. With this optimized system, we also expressed four GC-rich genes that are involved in the non-reversal peptide biosynthesis. Our results indicated that the percent solubility of all four proteins was notably higher as compared to the E. coli-based cell-free system, showing promise for soluble expression of high GC-continuous genes. In addition, we applied the cell-free system to other structurizing strains and found that all cell extracts of the test gene strains were active to produce the report protein. To summarize, in this work, we established a high-yielding crystallization-based cell-free protein synthesis system. Looking forward, we believe that our system will contribute significantly to express complex natural-product gene clusters, providing a new avenue for obtaining and studying natural-product biosynthesis pathways. With that, I would like to thank you for watching this video abstract. If you have any questions or comments, please feel free to contact us via email address that provided in the paper. Happy reading! Thank you!