Aspartokinase III, encoded by lysC, is responsible for the first step of lysine biosynthesis in Escherichia coli. In this study, a lysC knockout E. coli W3110 strain was generated to study the differential gene expression profiles of wild type and lysC knockout strains. Several significant changes were observed, including biosynthesis of lysine, oxaloacetate, alpha-ketoglutarate and glutamate genes. Genes related to transporters and heat shock proteins were also affected by lysC knockout. The results indicated that the lysC knockout strain exhibited some phenomena similar to lysine starvation. The data generated by this study further clarify the systematic role of lysC in lysine biosynthesis.