The invention discloses a method for screening a
corynebacterium crenatum endogenous high-expression
promoter by using a 2-DE (Two-Dimensional
Electrophoresis) technique, which belongs to the fields of
proteomics and
gene engineering. The method comprises the following steps of: identifying a high-
expression protein point in a
corynebacterium crenatum mycetocyte through the 2-DE technique in combination with a
mass-spectrometric technique; and
cloning encoding
gene promoters, naming as P-argC, P-argG, P-argF, P-ilvC and P-serA respectively, replacing a tac
promoter on a
shuttle vector pDXW-8, inserting a cat
gene downstream, transforming colon bacillus JM109 and
corynebacterium crenatum SYPA5-5 respectively, and measuring the activity of CAT (
Catalase), wherein the activities of the prompters P-argG, P-argF, P-ilvC, P-serA and P-argC in a recombinant strain decrease in sequence. According to the method, a constitutive high-expression
promoter from
corynebacterium crenatum is obtained for the first time, and an effective tool is provided for a
corynebacterium crenatum high-expression exogenous gene.