| 摘要 |
PURPOSE: An E.coli secreting human granulocyte-colony stimulating factor(hG-CSF) is provided, thereby secreting the hG-CSF effectively into cytoplasm and easily producing it through the simple processes. CONSTITUTION: A plasmid vector pTHKCSFmII comprises Kanamycin-tolerance gene, endozylanase secreting signal sequence, nucleotide sequence of SEQ ID NO: 1 encoding oligopeptide, human granulocyte-colony stimulating factor(hG-CSF) gene comprising the nucleotide sequence of SEQ ID NO: 26 encoding N-terminal, and Trc promoter, wherein the C-terminal of oligopeptide is Ile-Glu-Gly-Arg. E.coli MC4100/pTHKCSFmII (KCTC 0754BP) is transformed with plasmid vector pTHKCSFmII, wherein E.coli is selected from the group consisting of E.coli XL1-Blue, E.coli MC4100, E.coliBL21(DE3) , E.coli HB101 and E.coli W3110. The method for producing hG-CSF comprises the steps of: incubating E.coli MC4100/pTHKCSFmII (KCTC 0754BP) to produce hG-CSF fusion proteins; and treating hG-CSF with protease to produce hG-CSF.
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