| 摘要 |
PURPOSE: A method for mass-producing a recombinant human tyrosinase in E. coli is provided, thereby stably expressing the recombinant human tyrosinase having enzyme activity which is useful for studying on skin diseases from E. coli without forming inclusion body. CONSTITUTION: The method for mass-producing a recombinant human tyrosinase in E. coli comprises the steps of: (a) PCR amplifying the human tyrosinase gene having the nucleotide sequence set forth in SEQ IN NO: 1 using a pair of primers having the nucleotide sequences set forth in SEQ ID NO: 2 and SEQ ID NO: 3; (b) inserting the amplified human tyrosinase gene into a vector pET-26b(+) to prepare a recombinant expression vector pET-hTyr; (c) transforming E. coli with the recombinant expression vector pET-hTyr; and (d) culturing the transformed E. coli, isolating purifying the recombinant human tyrosinase, and subjecting it to DEAE=Sephacel anion exchange chromatography and His-Bind resin chromatography.
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