This invention discloses a method for preparing
inclusion bodies based on dual-
promoter expression plasmids and its application, belonging to the field of
biotechnology. The invention includes screening of
totipotent nucleases, purification of
totipotent nucleases,
verification of the degradation activity of the
totipotent nuclease DNS on nucleic acids, construction of dual-
promoter recombinant strains, fermenter culture,
cell recovery, and
cell disruption to obtain crude
inclusion bodies. This invention achieves high-efficiency expression of the recombinant
human growth hormone gene induced by the T7
promoter and high-activity expression of the totipotent
nuclease gene DNS induced by the cspA promoter, demonstrating significant effects in the application of recombinant
human growth hormone. During the preparation process, the
cell viscosity is low, the homogenization and disruption effect is improved, the BL21(DE3) / DGH
cell disruption rate is high, the inclusion body content harvested by
centrifugation per
unit volume of
fermentation broth is high, and the residual amount of HCD (host
DNA residue) in the
target protein per
unit mass is reduced, effectively reducing
energy consumption.