Background: The type VI secretion system (T6SS) is a recently described molecular machinery for protein transport across the cell envelope of a wide range Gram-negative bacteria, including many important human pathogens. The T6SS an attractive novel target for antibiotic development and such inhibitors will likely avoid the development of resistance. The applicant group leader (Sjöstedt A) has performed high throughput screening to identify the inhibitors of the VipA-VipB interaction of T6S using the bacterial two-hybrid interaction assay and few inhibitors were identified. These inhibitors will serve as blueprints for the identification of additional compounds with more potent inhibitory properties as well as optimal ADME profiling. The literature survey indicates that no drugs yet have been developed for T6SS targets. In this project, we are proposing advanced methodologies to identify inhibitors and antibacterial effector proteins of the T6SS.