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Title: Engineering by PCR-based exon amplification of the genomic porcine interferon-gamma DNA for expression in Escherichia coli
Authors: Vandenbroeck, K ×
Dijkmans, R
Van Aerschot, Arthur
Billiau, A #
Issue Date: Nov-1991
Series Title: Biochemical and Biophysical Research Communications vol:180 issue:3 pages:1408-15
Abstract: Interferon-gamma (IFN-gamma) is coded for by a single gene containing three introns, localized within the coding region. We have previously cloned the IFN-gamma gene from a pig genomic DNA lambda library and have determined its nucleotide sequence. In order to construct the porcine IFN-gamma DNA without intervening sequence, the four exons were separately amplified by the polymerase chain reaction (PCR) using primers matching the exon-termini. From the amplified exon-fragments the complete intron-free DNA was obtained by a strategy consisting of alternate rounds of PCR and ligation. The sequence so-obtained was used for expression in E. coli. The recombinant protein appeared as inclusion bodies which were solubilized and refolded in order to obtain biologically active IFN-gamma.
ISSN: 0006-291X
Publication status: published
KU Leuven publication type: IT
Appears in Collections:Medicinal Chemistry (Rega Institute)
Laboratory of Immunobiology (Rega Institute)
× corresponding author
# (joint) last author

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