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Hoffmann, R. A wiki for the life sciences where authorship matters. Nature Genetics (2008)
 

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Engineering by PCR-based exon amplification of the genomic porcine interferon-gamma DNA for expression in Escherichia coli.

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.[1]

References

  1. Engineering by PCR-based exon amplification of the genomic porcine interferon-gamma DNA for expression in Escherichia coli. Vandenbroeck, K., Dijkmans, R., van Aerschot, A., Billiau, A. Biochem. Biophys. Res. Commun. (1991) [Pubmed]
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