New bifunctional urokinase derivs and related plasmids

摘要

Bifunctional urokinase derivs. of formula M4-X1-Y1 (I) are new, where M4= sequence 47Ser to 411 Leu of the unglycosylated prourokinase sequence given in the fig.; X1=(Ser-Pro-Pro)2-A or Ser-X2-X3-X4-X5-X6-X7; A= Gly-Gly-Phe; Ser-Pro-Pro-Gly-Gly-Phe or Ser-Pro-Pro-Gly-Gly-Phe-Gly; X2 = Pro or Leu; X3 = Val or Pro; X4 = Lys, Val, Arg, Gly or Glu; X5 = Ala, Val, Gly, Leu or Ile; X6 = Phe, Trp, Tyr or Val; X7 = peptide bond or Gly; Y1 = Y2-Arg-Pro-Y3-(Gly)4-Asn-Gly-Asp-Phe-Glu-Glu-Ile-Pro-Glu- Glu-Tyr-Leu-Y4 (Y1a); Y2-Arg-Pro-Phe-Leu-Leu-Arg-Asn-Pro-Asn-Asp-Lys-Tyr-Glu-Pro- Phe-Trp-Glu-Asp-Glu-Glu-Lys-Asn-Glu (Y1b) or Y2-Arg-Pro-Ser-Ser-Glu-Phe-Glu-Glu-Phe-Glu-Ile-Asp-(Glu)3Lys; Y2 = Pro or Val; Y3= Leu or peptide bond; Y4= Gln or OH.Also new are plasmids, for expressing (I) in E.coli, in which the operon comprises a regulatable promoter; a Shine-Dalgarno sequence acting as ribosome binding site; a start codon; a synthetic structural gene for (I); and downstream of this 1 or 2 terminators.