METHOD FOR MANUFACTURING RECOMBINANT PROTEIN IN YEAST USING SECRETORY TYPE VECTOR

摘要

PURPOSE: A method for manufacturing a recombinant protein in yeast using a secretory type vector is provided, thereby resolving problems caused by using Escherichia coli and animal cells. CONSTITUTION: A method for manufacturing a recombinant human granulocyte-colony stimulating factor(hG-CSF) protein in yeast using a secretory type vector comprises using yeast transformed with a vector containing the nucleotide sequence encoding hG-CSF protein, wherein the yeast is Saccharomyces cerevisiae; the vector comprises the complex secretory signal sequence for increasing the secretory expression; the vector substitutes the yeast-favoring codons for a portion of the nucleotide sequence of yeast without changing the amino acid sequence in order to optimize the protein expression; the hG-CSF is expressed by a synthetic G-CSF gene containing the nucleotide sequence of SEQ ID NO: 46 substituted by the yeast-favoring codons; the vector comprises a ADH2/GAPDH mixed promoter, GAP termination sequence, marker gene, URA3 gene, and 2μ ori sequence; and the complex secretory signal sequence comprises the nucleotide sequence encoding the amino acid sequence of SEQ ID NO: 7.