Bacillus subtilis is gram-positive, rod-shaped, soil bacteria that became a model organism in many laboratory studies. Like many other bacteria, B. subtilis can divide by a very simple process of binary fission. It begins at middle of the cell, where cytoskeletal protein FtsZ forms a Z-ring that constricts progressively until the cell divides into two identical daughter cells. In conditions that are not sufficient enough to support vegetative growth, B. subtilis may undergo the simplest process of differentiation, called sporulation [2]. Unlike vegetative growth, sporulation septum is formed closer to one pole. After the axial filament has formed, the midcell Z-ring is converted into a spiral-like structure that travels towards each cell pole. The spiral then splits into two polar spirals and redeployment of FtsZ results in formation of two separate Z-rings near the two poles. This redeployment of the Z-ring requires SpoIIE, crucial sporulation protein. Finally, only one of two polar Z-rings becomes the site of asymmetric division [3]. Asymmetric septum forms around 1/6^th of the cell length and divides the cell into larger – mother cell and smaller – forespore. Asymmetric division involves almost the same set of proteins as vegetative division. However, several proteins play an important role in finding the site of asymmetric septum formation such as SpoIIE, MinCD and RefZ [1]. Here using bacterial two hybrid system (BACTH) and size exclusion chromatography (SEC), we searched for interaction partners of SpoIIE that might be involved in asymmetric septum formation. In addition, we followed localization and co-localization of these proteins by super resolution microscopy (SIM).