國家衛生研究院 NHRI:Item 3990099045/1145
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    题名: Sequential expression of recombinant proteins and their separate recovery from a Pichia pastoris cultivation
    作者: Wu, JM;Chieng, LL;Hsu, TA;Lee, CK
    贡献者: Division of Biotechnology and Pharmaceutical Research
    摘要: Pichia pastoris strains with capability to sequentially express two recombinant proteins under control of the constitutive glyceraldehydes-3-phosphate dehydrogenase (GAP) and the methanol-induced alcohol oxidase (AOX1) promoters have been constructed. The intracellular beta-galactosidase (beta-gal), secreted human serum albumin (hSA) and human granulocyte-macrophage colony-stimulating factor (hGM-CSF) are used as model proteins to demonstrate their sequential expression and separate recovery. The strain X33/ZCSF cultivated in YPD medium results in high-level constitutive expression of p-galactosidase activity and methanol-induced, secreted highly glycosylated hGM-CSF expression. The p-galactosidase activity and hGM-CSF are separately recovered from cells pellet and supernatant. On the other hand, the strain GS115/CSFA cultivated in YPD medium constitutively expresses secreted hGM-CSF along with cell growth. After a high cell concentration is achieved, the YPD medium is then replaced with YPM medium to induce the expression of the AOX1 promoter controlled hSA. The sequentially expressed hGM-CSF and hSA can be separately recovered from the spent YPD and YPM medium. (C) 2003 Elsevier Science B.V. All rights reserved.
    关键词: Biotechnology & Applied Microbiology;Engineering, Chemical
    日期: 2003-10
    關聯: Biochemical Engineering Journal. 2003 Oct;16(1):9-16.
    Link to: http://dx.doi.org/10.1016/S1369-703X(03)00017-2
    JIF/Ranking 2023: http://gateway.webofknowledge.com/gateway/Gateway.cgi?GWVersion=2&SrcAuth=NHRI&SrcApp=NHRI_IR&KeyISSN=1369-703X&DestApp=IC2JCR
    Cited Times(WOS): https://www.webofscience.com/wos/woscc/full-record/WOS:000185127000002
    Cited Times(Scopus): http://www.scopus.com/inward/record.url?partnerID=HzOxMe3b&scp=0041363483
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