Expression of a β-Lactamase Preproinsulin Fusion Protein in Escherichia Coli View Full Text


Ontology type: schema:ScholarlyArticle     


Article Info

DATE

1984-02

AUTHORS

Anne W. Emerick, Barbara L. Bertolani, A. Ben-Bassat, Thomas J. White, Michael W. Konrad

ABSTRACT

The level of expression of a β-lactamase human preproinsulin fusion protein1 in Escherichia coli was increased by plasmid and host manipulations. The initial expression level (0.01% of total protein) was increased two- to three-fold by replacing the p-lactamase promoter with two strong promoters: trp from Serratia marcescens and lacUV5 from E. coli. Replacement of the β-lactamase ribosome binding site (RBS) with a trp RBS enhanced the yield to a total of 0.16% of the total cell protein. Proinsulin was unstable, with a half-life of 14 minutes at 37°C. Proinsulin expression was increased three-fold (to about 0.5% of the total cell protein) by incubating the host at 30°C rather than 37°C. More... »

PAGES

165

Identifiers

URI

http://scigraph.springernature.com/pub.10.1038/nbt0284-165

DOI

http://dx.doi.org/10.1038/nbt0284-165

DIMENSIONS

https://app.dimensions.ai/details/publication/pub.1012678702


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