| 摘要 |
PURPOSE: A method for producing restriction enzyme in E.coli is provided to produce restriction enzyme with high stability and high efficiency using diluted culture medium. CONSTITUTION: A method for producing recombinant endonuclease in E.coli comprises: a step of transforming E.coli with a plasmid containing a gene encoding a restriction enzyme; a step of comparing transformed E.coli with standard concentration; a step of culturing the E.coli in diluted culture medium for 12 to 20 hours; a step of adding medium ingredient to be standard concentration; a step of adding inducer; and a step of culturing for 2-18 hours. The E.coli is BL21(DE3)/pLysS, Rosetta(DE3)/pLysS, Rosetta2(DE3)/pLysS, ArcticExpress(DE3), C41(DE3), or C43(DE3). The restriction enzyme is X or KII derived from Xanthomonas oryzae pv. Oryzae. The plasmid is pET28a/XorKII. |
