Cell-free synthesis of actin-like cytoskeleton filaments
MreB, an actin-like cytoskeleton in E. coli encoded by MreB gene, is essential for the maintenance of bacterial cell shape. Our project is aimed at the cell-free synthesis of MreB, and the actin homolog MreB was further characterized through both microfluidics technology and nanobiotechnology. As in vivo, MreB is rigidly linked to the cell wall. Here, MreB was cell-free expressed in vitro. Protein localization on the inner oil film could be observed in the W/O/W microdroplets produced with microfluidic technologies, which mimics the cell membrane to provide a non-polar environment. Furthermore, CNTs (carbon nanotubes) were involved as scaffolds for the auxiliary expression of actin homolog MreB. The combination of MreB with CNTs and protein aggregation were observed though the AFM (atomic force microscope) in vitro. Our research cell-free synthesizes the actin-like cytoskeleton filaments, and provides a sound microenvironment for protein synthesis, which helps to recognize, explore and create life.