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- W2371825419 abstract "Objective To generate two mutants of human soluble APRIL(sAPRIL) for laying a foundation for developing competitive inhibitor of a proliferation inducing ligand(APRIL). Methods Taking 186G and 187Q of human APRIL as mutation sites, one-step opposite orientation PCR was used to construct the two mutant sAPRIL(msAPRIL) DNAs, namely msAPRIL-1 DNA(in which 186G was replaced by K while 187Q was deleted) and msAPRIL-2 DNA(in which 186G187Q was replaced by 186K187G). The two sequenced msAPRIL DNAs were separately subcloned into the prokaryotic expression vector pQE-80L, and the corresponding recombinant msAPRIL proteins were expressed in E. coli DH5α. The expressed msAPRIL proteins were analyzed by SDS-PAGE and Western blotting, and purified by Ni2+-NTA and Sephadex G-75 chromatography. Results By using one-step opposite orientation PCR, the two msAPRIL DNAs were cloned, and their sequences were consistent with the designed mutation. After inserting every msAPRIL DNA into pQE-80L vector respectively, the recombinant msAPRIL proteins were successfully expressed in E. coli DH5α. The msAPRIL proteins were purified effectively; the homotrimers of msAPRILs were isolated by Sephadex G-75. Conclusion Two msAPRIL proteins are successfully prepared, which might pave a way for developing novel antitumor therapeutics based on mutant sAPRIL." @default.
- W2371825419 created "2016-06-24" @default.
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- W2371825419 date "2007-01-01" @default.
- W2371825419 modified "2023-09-23" @default.
- W2371825419 title "Prokaryotic expression and purification of human soluble APRIL mutants" @default.
- W2371825419 hasPublicationYear "2007" @default.
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