The cell envelope of Gram-negative bacteria is typically composed of the inner and outer membranes (IM and OM, respectively), an aqueous compartment known as the periplasm, and a mesh-like layer known as peptidoglycan (PG). Proper envelope biogenesis is crucial for the bacterium and it requires the coordinated synthesis, transport and assemblage of all its components. In our laboratory, we use genetic and biochemical approaches to understand envelope biogenesis in the Gram-negative bacterium Escherichia coli.