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- W2393217878 abstract "Objective:To construct eukaryotic expression vector of ITF and express the recombinant ITF in Pichia pastoris GS115.The research establish the base for further research of physiological and pharmaceutical functions of ITF.Methods:ITF DNA fragment was amplified using polymerase chain reaction (PCR),the gene was doned into the yeast expression vector pPICZαA,and the recombinant expression vector pPICZαA-ITF gained.The recombinant plasmid pPICZαA-ITF was linearized by BspH I and transformed into the yeast strain GS115 with lithium chloride.Zeocin resistant transformed yeast strains were cho- sen,and the presence of insert was identified by PCR.The positive transformants were expressed and the proteins in the culture supernatant were deposited in flask with TCA and analyzed with Tricine SDS-PAGE and Western blotting.Result:PCR result and gene sequencing proved that the fragment amplified was inserted into the yeast expression vector pPICZαA correctly.It was proved that the molecular weight of expressed proteins was about 14×10~3 by Tricine SDS-PAGE analysis and Western blotting showed the proteins have good antigenicity and specificity.Conclusions:pPICZαA-ITF eukaryotic expression vector was con- structed successfully,recombinant ITF was expressed in yeast fungus GS115 and recombinant ITF protein was gained." @default.
- W2393217878 created "2016-06-24" @default.
- W2393217878 creator A5032820759 @default.
- W2393217878 date "2007-01-01" @default.
- W2393217878 modified "2023-09-23" @default.
- W2393217878 title "Construction of Intestinal Trefoil Factor Expression Vector and Expression of ITF in Yeast Fungus GS115" @default.
- W2393217878 hasPublicationYear "2007" @default.
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