SEAFDEC/AQDINSTITUTIONAL REPOSITORY
    • Login
    View Item 
    •   SEAFDEC/AQD Institutional Repository Home
    • 03 SEAFDEC/AQD External Publications
    • Journal Articles, Conference Papers and Book Chapters by SEAFDEC Staff
    • Journal Articles
    • View Item
    •   SEAFDEC/AQD Institutional Repository Home
    • 03 SEAFDEC/AQD External Publications
    • Journal Articles, Conference Papers and Book Chapters by SEAFDEC Staff
    • Journal Articles
    • View Item
    JavaScript is disabled for your browser. Some features of this site may not work without it.

    Expression and purification of a biologically active recombinant rabbitfish (Siganus guttatus) growth hormone

    • Global styles
    • APA
    • Elsevier - Harvard
    • Vancouver
    • MLA
    • Help
    Thumbnail
    View/Open
    Request a copy
    Date
    2005
    Author
    Funkenstein, Bruria
    Dyman, Dyman
    Lapidot, Ziva
    de Jesus-Ayson, Evelyn Grace
    Gertler, Arieh
    Ayson, Felix G.
    Page views
    753
    Metadata
    Show full item record

    Cited times in Scopus

    
    
    Share 
     
    Abstract
    Recombinant rabbitfish growth hormone (rfGH) protein was expressed in Escherichia coli, BL21(DE3) cells. The cDNA encoding the mature protein of rfGH was first cloned in pGEM-Teasy vector and then transferred to pET-3d expression vector. Expression in E. coli cells was then induced by IPTG (0.4 mM). Inclusion bodies (IB) containing the expressed protein were purified by treating bacterial cells pellet with lysozyme followed by repeated washings in cold water containing Triton X-100, sonication, and centrifugation. IB were then solubilized in 4.5 M urea, refolded at pH 11.3 in the presence of catalytic amounts of cysteine and purified by Q-Sepharose column. Gel filtration on Superdex column showed the purified protein to be a monomeric GH. Based on SDS–PAGE, the purity of the recombinant rfGH preparation is approximately 98%. The recombinant rfGH was tested for its biological activity both in vitro, by its ability to stimulate IGF-I mRNA expression in the liver, and in vivo, by its ability to accelerate growth in rabbitfish fry injected with the hormone. A significant increase in growth was observed in rabbitfish fry given the recombinant hormone. Polyclonal antibody raised against the native rfGH immunoreacted with the recombinant rfGH in Western blots and in ELISA, indicating the suitability of these reagents for future quantification of GH in rabbitfish plasma.
    URI
    http://hdl.handle.net/10862/2032
    Suggested Citation
    Funkenstein, B., Dyman, D., Lapidot, Z., de Jesus-Ayson, E. G., Gertler, A., & Ayson, F. G. (2005). Expression and purification of a biologically active recombinant rabbitfish (Siganus guttatus) growth hormone. Aquaculture, 250(1-2), 504-515.
    DOI
    10.1016/j.aquaculture.2005.04.065
    Type
    Article
    ISSN
    0044-8486
    Subject
    Antibodies; Bacterial diseases; Centrifugation; Cysteine; Disease transmission; Elisa; Filtration; Fish culture; Fish diseases; Gene expression; Growth; Hormones; Husbandry diseases; Liver; Marine fish; Recombinants; Urea; pH effects; Expression vectors; Growth hormone; Inclusion bodies; Insulin-like growth factor I; Lysozyme; Escherichia coli; Siganus guttatus; Israel
    Collections
    • Journal Articles [1100]


    © SEAFDEC/AQD  2021
    Contact Us | Send Feedback
     

     

    Browse

    All of SAIRCommunities & CollectionsBy Issue DateAuthorsTitlesSubjectsKeywordsThis CollectionBy Issue DateAuthorsTitlesSubjectsKeywords

    My Account

    LoginRegister

    Statistics

    View Usage Statistics

    © SEAFDEC/AQD  2021
    Contact Us | Send Feedback
     

     

    Export citations

    Export the current results of the search query as a citation list. Select one of the available citation styles, or add a new one using the "Citations format" option present in the "My account" section.

    The list of citations that can be exported is limited to items.

    Export citations

    Export the current item as a citation. Select one of the available citation styles, or add a new one using the "Citations format" option present in the "My account" section.

    Export Citations

    EXTERNAL LINKS DISCLAIMER

    This link is being provided as a convenience and for informational purposes only. SEAFDEC/AQD bears no responsibility for the accuracy, legality or content of the external site or for that of subsequent links. Contact the external site for answers to questions regarding its content.

    If you come across any external links that don't work, we would be grateful if you could report them to the repository administrators.

    Click DOWNLOAD to open/view the file. Request a copy in case the link we provided don't work.

    Download

    DOCUMENT REQUEST NOT AVAILABLE

    This publication is still available (in PRINT) and for sale at AQD bookstore. The library is currently restricted to send PDF of publications that are still for sale.

    You may contact bookstore@seafdec.org.ph or visit AQD bookstore for orders.

    FILE UNDER EMBARGO

    This file associated with this publication is currently under embargo. This will be available for download after the embargo date.