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- W2373671510 abstract "Objective:To study the construction and expression of mature peptide vector of human osteoprotegerin(OPG)and the immunoassay of its recombinant protein.Methods:Total RNA was extracted from primary human osteoblasts.The cDNA encoding the mature peptide of OPG was amplified through RT-PCR and inserted into the secretory expression vector pPIC9K.The recombinant plasmid was then transformed into GS115,a pichia strain.Mut+ phenotypic transformants were selected according to their growth on MM and MD plates.These colonies were further screened for transformants with multicopy inserts integretion by evaluating their growth on YPD plates with different concentration of G418.Finally,the G418-resistant colonies were cultured in BMMY medium at 30 ℃ in a shaking incubator under the induction of 0.5% methanol.Results:The recombinant plasmid was successfully constructed.A transformant resistant to 3.0 g/L G418 was screened out for expression.Induced with methanol,the cells secreted a 55_ku protein that represented about 30% of total protein in the supernatant on SDS-PAGE gel.The 55_ku protein could be recognized,through Western blotting analysis,by the polyclonal antibody against OPG.Conclusion:The construction and expression of the recombinant vector for mature peptide of OPG is of much value since it provides a sound basis for further research on ODF/OPG/PANK system and a promising pharmaceutical tool for osteoporotic and erosive bone disorders." @default.
- W2373671510 created "2016-06-24" @default.
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- W2373671510 date "2003-01-01" @default.
- W2373671510 modified "2023-09-23" @default.
- W2373671510 title "Construction and Expression of Recombinant Vector for Mature Peptide of Human Osteoprotegerin and Immunoassay of Its Recombinant Protein" @default.
- W2373671510 hasPublicationYear "2003" @default.
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