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- W2380973989 abstract "Objective To design and clone full human cardiac troponin I (hcTnI) gene and construct an efficient expression system for hcTnI in prokaryotic cells. Methods According to the codon usage bias of E.coli, an artificial gene was designed for cTnI. The gene was synthesized, ligated by PCR, and then cloned into a prokaryotic nonfusion-protein type vector pBV220 and subsequently transformed into E.(coli) DH5α, so as to construct prokaryotic nonfusion-protein expression bacterial hcTnI- pBV220/DH5α. After heat shock, the hcTnI nonfusionprotein was induced to express, and then identified by SDS-PAGE. The expressed hcTnI nonfusion protein was the specific antigen against anti-(cTnI) monoclonal antibodies. Results The nucleotide sequence analysis indicated that the sequence of cloned artificial cTnI gene was accorded with that of designed. The positive clones were primarily confirmed by anti-ampicillin selection and restrictive enzyme digestion. The relative molecular weight of hcTnI nonfusionprotein was about M_r 24 000 identified by SDS-PAGE The active human cTnI protein was obtained and amounted to 25 % of total bacterial protein. Conclusion The human cTnI artificial gene is obtained and successfully constructed its prokaryo-(tic) expression vector, which will provide theoretical basis for preparation of high-specific monoclonal antibody and clinical laboratory diagosis of cTnI." @default.
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- W2380973989 date "2005-01-01" @default.
- W2380973989 modified "2023-09-23" @default.
- W2380973989 title "Cloning and overexpression of human cardiac troponin I gene" @default.
- W2380973989 hasPublicationYear "2005" @default.
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