Secretion of human EGF and IgE in mammalian cells by recombinant DNA techniques; use of a IL-2 leader sequence.
Expression plasmids were constructed containing chemically synthesized human epidermal growth factor (EGF) gene fused in a frame to a leader sequence of human interleukin-2 (IL-2) gene under the control of a viral promoter. COS7 cells transfected with the plasmids synthesized and secreted EGF. Transfection of mouse A9 cells or BALB/3T3 clone A31 cells with the plasmids permitted the isolation of cell lines secreting the product which showed EGF activity. In particular, A31 transformed cells secreting human EGF grew well even in a medium containing a minimal level of serum. Using similar vectors having IgE cDNA (C2-C4) in place of EGF gene, a human IgE Fc fragment was also produced and secreted in mouse cells. These results show that heterologous leader sequences are useful for the expression and secretion of proteins whose genes lack leader sequences.
Sasada
R
R
Central Research Division, Takeda Chemical Industries, Ltd., Osaka, Japan.
Marumoto
R
R
Igarashi
K
K
D000818:Animals; D001483:Base Sequence; D002460:Cell Line; D004274:DNA, Recombinant; D004815:Epidermal Growth Factor / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D007073:Immunoglobulin E / Q000032:analysis / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism*; D007376:Interleukin-2 / Q000235:genetics*; D051379:Mice; D008969:Molecular Sequence Data; D010957:Plasmids; D021382:Protein Sorting Signals / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis / Q000235:genetics
D004274:DNA, Recombinant; D007376:Interleukin-2; D021382:Protein Sorting Signals; D011993:Recombinant Fusion Proteins; D007073:Immunoglobulin E; D004815:Epidermal Growth Factor