Organelles are connected and exchange proteins and lipids each other by vesicular transport, while maintaining their specific morphology and components during interphase. At the onset of mitosis, the balance of vesicular transport between organelles is disturbed, resulting in the drastic change of organelle morphology. To understand the coupling mechanisms between organelle morphology and vesicular transport, we developed the reconstitution systems for the Golgi disassembly, the disruption and reformation of the endoplasmic reticulum (ER), mitotic disruption of ER exit sites, and vesicular transport between ER and Golgi in semi-intact cells using interphase or mitotic cytosol, and elucidated biochemical requirements for the processes. In this paper, we will show the molecular links between organelle morphology and transport, especially focused on the mitotic kinases and its regulation of membrane fusion machineries.