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- W2347735404 abstract "Objective To construct eukaryotic expression plasmid pEE14.1-dsFvαpr+,and detect the expression of the recombined gene in eukaryotic CHO-K1 cells.Methods The cationic DNA fragment was cloned into the 3' of VH gene by overlapping extension PCR,and the 6×His tab was inserted to the 3' of VL and human IFN-α gene by the same way.The above mentioned recombinant VH and VL genes were inserted into a pCI-GPI vector first,and then cloned into the pEE14.1 vector to construct the recombinant plasmid pEE14.1-dsFvαpr+.Finally,the recombinant plasmid was transfected into the CHO-K1 cells by LipofectamineTM 2000,and the expression was detected by RT-PCR,ELISA and Western blotting.Results The enzyme digestion and sequencing analysis showed that the recombinant plasmid was successfully constructed.RT-PCR showed that only the cells with transfected plasmid can generate the specific 1700bp fragment.ELISA analysis showed that the production of IFN-αexpressed in the supernatant of transfected cells was about 1.1ng/ml.Also,Western blotting could reveal the characteristic band of HBsAg dsFvαpr+ protein.Conclusion The antibody targeting to human IFN-αgenes has been successfully expressed in a single open reading frame.Changing the electricity of the antibody may provide the necessary condition for the study of the a new type of anti-HBV drug in nanoscale in the future." @default.
- W2347735404 created "2016-06-24" @default.
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- W2347735404 date "2010-01-01" @default.
- W2347735404 modified "2023-09-22" @default.
- W2347735404 title "Construction and expression of human anti-HBs-IFN fusion gene" @default.
- W2347735404 hasPublicationYear "2010" @default.
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