University of Venda 1 article published in JoVE Biochemistry Escherichia coli -Based Complementation Assay to Study the Chaperone Function of Heat Shock Protein 70 Hloniphani Rachel Ncube1, Unarine Dali1, Tendamudzimu Harmfree Dongola1, Addmore Shonhai1 1Department of Biochemistry & Microbiology, University of Venda This protocol demonstrates the chaperone activity of heat shock protein 70 (Hsp70). E. coli dnaK756 cells serve as a model for the assay as they harbor a native, functionally impaired Hsp70, making them susceptible to heat stress. The heterologous introduction of functional Hsp70 rescues the growth deficiency of the cells.