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- W2388066713 abstract "Objective To e xp ress thyroid hormone-response protein-1 (TRP-1) by gene engeneering technique . Methods The gene encoding TRP-1 of RNA from neonatal rat brain tissue was amplified by RT-PCR, and was subcloned into Pinpo int Xa-1 T vector to construct recombinant expression plasmid. After confirming the sequence and open reading frame by DNA sequencing, the expression of target protein in E. coli was induced by isopropyl β-D-thiogalacto pyranoside (IPTG ). The fusion protein was detected by Western blotting. Using avidin resin, the biotinylated fusion protein was purified under non-denaturing condition by affi nity chromatograph and its molecutar weight was measured by SDS-PAGE. Results The specific PCR products were correctly inserte d into restriction sites of pinpoint Xa-1 expression vector. The result of West ern blotting indicated that the fusion protein was expressed as expected, the fu sion protein was purified by affinity chromatograph, and SDS-PAGE showed that t he molecular weight of the TRP-1 fusion protein was 23400 as ca lculated according to its amino sequence. Conclusion Using prokaryocyte expression system, TRP-1 fusion protein is expressed, thus, it is possible to further investigate the function of this protein and its role in brain development and other tissues." @default.
- W2388066713 created "2016-06-24" @default.
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- W2388066713 date "2003-01-01" @default.
- W2388066713 modified "2023-09-25" @default.
- W2388066713 title "Prokaryotic expression of thyroid hormone-response protein-1 gene" @default.
- W2388066713 hasPublicationYear "2003" @default.
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