The invention discloses a method for labeling escherichia coli proteome by using SILAC (Stable Isotope Labeling with Amino Acids in Cell Cultures) and a special culture medium. The invention provides a special culture medium for the SILAC-labeled escherichia coli. The special culture medium comprises inorganic salt, histidine hydrochloride, isoleucine, valine, leucine, phenylalanine, tryptophan, serine, heavy stable isotope labeled lysine, arginine, threonine, tyrosine, methionine, adenine sulfate, uracil, vitamin B1 and glucose; and the inorganic salt is Na2HPO4.12H2O, KH2PO4, NaCl, MgSO4, NH4Cl and CaCl2. Experiments prove that, the special culture medium for the SILAC-labeled escherichia coli, disclosed by the invention, creates a condition for researches of labeling the escherichia coli proteome, preparing a quantitative internal label and highly-precisely quantifying the escherichia coli proteome.