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Please use this identifier to cite or link to this item: http://repository.li.mahidol.ac.th/dspace/handle/123456789/21212
Title: Expression of human pyruvate carboxylase in insect cells using the baculovirus system
Authors: Natthida Petchampai
Vichai Boonsaeng
John C. Wallace
Sarawut Jitrapakdee
Mahidol University
University of Adelaide
Keywords: Biochemistry, Genetics and Molecular Biology
Issue Date: 26-Mar-2004
Citation: Biochemical and Biophysical Research Communications. Vol.316, No.1 (2004), 177-181
Abstract: Constructs containing cDNA encoding human pyruvate carboxylase (PC) with and without a hexahistidine (6x His) tag at the N-terminal of the mature enzyme have been cloned under the control of the polyhedrin promoter. These two constructs were co-transfected with the baculovirus genome into Sf9 cells to produce recombinant baculoviruses harbouring human PC cDNA. The expression of human PC under the control of the polyhedrin promoter was found to be at its highest level at 4 days post-infection. The expressed material accounted for up to 70% of total cellular protein with 5% of this expressed material being found in the soluble fraction. The recombinant human 6x His-PC isolated with a purity of ∼50% using a Ni-NTA agarose column was found to have the specific activity of 7U/mg, which was similar to that produced from a 293T stable line [Biochem. Biophys. Res. Commun. 266 (1999) 512]. This is the first report of a heterologous expression system for recombinant human PC. © 2004 Elsevier Inc. All rights reserved.
URI: https://www.scopus.com/inward/record.uri?partnerID=HzOxMe3b&scp=1542345765&origin=inward
http://repository.li.mahidol.ac.th/dspace/handle/123456789/21212
ISSN: 0006291X
Appears in Collections:Scopus 2001-2005

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