A genomically modified Escherichia coli strain carrying an orthogonal E. coli histidyl-tRNA synthetase•tRNA^His pair.

ABSTRACT:

Background

Development of new aminoacyl-tRNA synthetase (aaRS)•tRNA pairs is central for incorporation of novel non-canonical amino acids (ncAAs) into proteins via genetic code expansion (GCE). The Escherichia coli and Caulobacter crescentus histidyl-tRNA synthetases (HisRS) evolved divergent mechanisms of tRNA^His recognition that prevent their cross-reactivity. Although the E. coli HisRS•tRNA^His pair is a good candidate for GCE, its use in C. crescentus is limited by the lack of established genetic selection methods and by the low transformation efficiency of C. crescentus.

Methods

E. coli was genetically engineered to use a C. crescentus HisRS•tRNA^His pair. Super-folder green fluorescent protein (sfGFP) and chloramphenicol acetyltransferase

SUBMITTER: Englert M 

PROVIDER: S-EPMC5592127 | biostudies-literature | 2017 Nov

REPOSITORIES: biostudies-literature