A mutant library of subtilisin E containing random combinations of various mutagenized sites wasconstructed by one-round mutagenesis with 15 mutagenic oligonucleotides. Mutants were screened through dot blot hybridiza...A mutant library of subtilisin E containing random combinations of various mutagenized sites wasconstructed by one-round mutagenesis with 15 mutagenic oligonucleotides. Mutants were screened through dot blot hybridization and DNA sequencing. A single-point mutant (Met 222Ala) and a three-point (Asn 76Asp/Asnl09Ser/ I le 205/Cys) mutant gene from the library were expressed. The mutant proteins exhibited conspicuously improved resistance to oxidation and heat treatment, as reported before. The results show that the library is reliable and very useful for protease subtilisin E engineering.展开更多
Pro-urokinase is a kind of protease existing in human body. It has the character of fibrinolysis, so it has been clinically applied to the treatment of thrombolytic diseases. In order to produce a large amount of pro-...Pro-urokinase is a kind of protease existing in human body. It has the character of fibrinolysis, so it has been clinically applied to the treatment of thrombolytic diseases. In order to produce a large amount of pro-urokinase by genetic engineering method and reform it with protein engineering, we have chemically synthesized the pro-urokinase gene, which展开更多
基金Project supported by United Nations Industrial Development Organization (UNIDO).
文摘A mutant library of subtilisin E containing random combinations of various mutagenized sites wasconstructed by one-round mutagenesis with 15 mutagenic oligonucleotides. Mutants were screened through dot blot hybridization and DNA sequencing. A single-point mutant (Met 222Ala) and a three-point (Asn 76Asp/Asnl09Ser/ I le 205/Cys) mutant gene from the library were expressed. The mutant proteins exhibited conspicuously improved resistance to oxidation and heat treatment, as reported before. The results show that the library is reliable and very useful for protease subtilisin E engineering.
文摘Pro-urokinase is a kind of protease existing in human body. It has the character of fibrinolysis, so it has been clinically applied to the treatment of thrombolytic diseases. In order to produce a large amount of pro-urokinase by genetic engineering method and reform it with protein engineering, we have chemically synthesized the pro-urokinase gene, which
