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- W2369546957 abstract "Objective:To provide reliable and effective method of getting activated human caspase-3. Methods: The cDNA of procaspase-3 was subcloned into E coli expression vector pMTY4-His. MS2-procaspase-3 fusion protein was expressed in E coli as inclusion bodies (IB). The IB were solubilized in guanidine hydrochloride and dialyzed against 50% acetic acid overnight and diluted with 9 volumes of water. The mixture was diluted rapidly to one containing 1.0 mol·L -1 Tris, pH 8.0 and 5 mmol·L -1 DTT and then concentrated to a nearly-dry state on the ultrafiltration membrane, followed by dissolving the concentrated protein. Results: High level expression of MS2-procaspase-3 recombinant protein was achieved in E coli as IB. About 10 mg activated caspase-3 could be obtained from 1L cultured bacteria. The activated caspase-3 could efficiently cleave Ac-DEVD-AMC and was inhibited by Ac-DEVD-CHO. Conclusion: We have successfully established a procedure for obtaining large quantity of activated caspase-3,which could be used for studying the function and structure of caspase-3." @default.
- W2369546957 created "2016-06-24" @default.
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- W2369546957 date "2002-01-01" @default.
- W2369546957 modified "2023-09-25" @default.
- W2369546957 title "Over-expression and autoactivation of recombinant human procaspase-3" @default.
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