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

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Author list: Jennings MJ, Barrios AF, Tan S
Publisher: Elsevier
Publication year: 2016
Volume number: 121
Start page: 17
End page: 21
Number of pages: 5
ISSN: 1046-5928
Languages: English-Great Britain (EN-GB)


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. (C) 2015 Elsevier Inc. All rights reserved.


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Last updated on 2019-23-08 at 11:15