FIELD: biotechnology. SUBSTANCE: invention relates to preparing the recombinant plasmid DNA "pДТИ-23" determining synthesis of the fused protein diphtherin toxin/interleukin-2 (DTI-2) and strain of bacterium Escherichia coli containing the indicated plasmid DNA. Size of recombinant plasmid DNA "pДТИ-23" is 4.7 thousand base pairs. Gene diphtheric toxin (DT) containing in this plasmid is under control of inducible lactose operon promoter from plasmid pV18. Method of preparing plasmid "pДТИ-23" involves linkage of frame reading of gene diphtheric toxin from corynephage ω and natural gene of the human mature interleukin-2 using Eco RI-site of gene DT, insertion of Bam HI-linker, treatment with restriction endonuclease Bam HI followed by ligation of DNA-fragment in vector plasmid. Strain E. coli grows well on the simple nutrient media, shows resistance to ampicillin and produces the fused protein DTI-2 after induction with isopropyl-b-D-thiogalactoside. Recombinant plasmid can be used in medicine and biological investigations of T-cells. EFFECT: improved method of plasmid preparing. 3 cl, 5 dwg
Authors
Dates
1997-04-20—Published
1993-12-10—Filed