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Protein Expr Purif. 2016 May;121:17-21. doi: 10.1016/j.pep.2015.12.001. Epub 2015 Dec 29.

Elimination of truncated recombinant protein expressed in Escherichia coli by removing cryptic translation initiation site.

Author information

1
Center for Eukaryotic Gene Regulation, Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA 16802, USA.
2
Center for Eukaryotic Gene Regulation, Department of Biochemistry and Molecular Biology, The Pennsylvania State University, University Park, PA 16802, USA. Electronic address: sxt30@psu.edu.

Abstract

Undesirable truncated recombinant protein products pose a special expression and purification challenge because such products often share similar chromatographic properties as the desired full length protein. We describe here our observation of both full length and a truncated form of a yeast protein (Gcn5) expressed in Escherichia coli, and the reduction or elimination of the truncated form by mutating a cryptic Shine-Dalgarno or START codon within the Gcn5 coding region. Unsuccessful attempts to engineer in a cryptic translation initiation site into other recombinant proteins suggest that cryptic Shine-Dalgarno or START codon sequences are necessary but not sufficient for cryptic translation in E. coli.

KEYWORDS:

Cryptic initiation; Escherichia coli expression; Recombinant protein expression

PMID:
26739786
PMCID:
PMC4803570
DOI:
10.1016/j.pep.2015.12.001
[Indexed for MEDLINE]
Free PMC Article

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