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

The level of expression of a β-lactamase human preproinsulin fusion protein 1 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: t...

Ausführliche Beschreibung

Gespeichert in:
Bibliographische Detailangaben
Veröffentlicht in:Bio/Technology 1984-02, Vol.2 (2), p.165-168
Hauptverfasser: Emerick, Anne W, Bertolani, Barbara L, Ben-Bassat, A, White, Thomas J, Konrad, Michael W
Format: Artikel
Sprache:eng
Schlagworte:
Online-Zugang:Volltext
Tags: Tag hinzufügen
Keine Tags, Fügen Sie den ersten Tag hinzu!
Beschreibung
Zusammenfassung:The level of expression of a β-lactamase human preproinsulin fusion protein 1 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 lac UV5 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.
ISSN:0733-222X
1087-0156
2331-3684
1546-1696
DOI:10.1038/nbt0284-165