Enhancing the Translational Capacity of E. coli by Resolving the Codon Bias

Result code in IS VaVaI

<a href="https://www.isvavai.cz/riv?ss=detail&h=RIV%2F00064190%3A_____%2F18%3AN0000111" target="_blank" >RIV/00064190:_____/18:N0000111 - isvavai.cz</a>

Result on the web

<a href="http://dx.doi.org/10.1021/acssynbio.8b00332" target="_blank" >http://dx.doi.org/10.1021/acssynbio.8b00332</a>

DOI - Digital Object Identifier

<a href="http://dx.doi.org/10.1021/acssynbio.8b00332" target="_blank" >10.1021/acssynbio.8b00332</a>

Result language

angličtina

Original language name

Enhancing the Translational Capacity of E. coli by Resolving the Codon Bias

Original language description

Escherichia coli is a well-established and popular host for heterologous expression of proteins. The preference in the choice of synonymous codons (codon bias), however, might differ for the host and the original source of the recombinant protein, constituting a potential bottleneck in production. Codon choice affects the efficiency of translation by a complex and poorly understood mechanism. The availability of certain tRNA species is one of the factors that may curtail the capacity of translation. Here we provide a tRNA-overexpressing strategy that allows the resolution of the codon bias, and boosts the translational capacity of the popular host BL21(DE3) when rare codons are encountered. In the BL21(DE3)-derived strain, called SixPack, copies of the genes corresponding to the six least abundant tRNA species have been assembled in a synthetic fragment and inserted into a rRNA operon. This arrangement, while not interfering with the growth properties of the new strain, allows dynamic control of the transcription of the extra tRNA genes, providing significantly elevated levels of the rare tRNAs in the exponential growth phase. Results from expression assays of a panel of recombinant proteins of diverse origin and codon composition showed that the performance of SixPack surpassed that of the parental BL21(DE3) or a related strain equipped with a rare tRNA-expressing plasmid.

Czech name

—

Czech description

—

Type

J_ost - Miscellaneous article in a specialist periodical

CEP classification

—

OECD FORD branch

30203 - Respiratory systems

Project

—

Continuities

V - Vyzkumna aktivita podporovana z jinych verejnych zdroju

Publication year

2018

Confidentiality

S - Úplné a pravdivé údaje o projektu nepodléhají ochraně podle zvláštních právních předpisů

Name of the periodical

ACS synthetic biology

ISSN

2161-5063

e-ISSN

2161-5063

Volume of the periodical

7

Issue of the periodical within the volume

11

Country of publishing house

US - UNITED STATES

Number of pages

8

Pages from-to

2656-2664

UT code for WoS article

—

EID of the result in the Scopus database

—