Fluorescence bimolecular complementation enables facile detection of ribosome assembly defects in Escherichia coli.

ABSTRACT: Assembly factors promote the otherwise non-spontaneous maturation of ribosome under physiological conditions inside the cell. Systematic identification and characterization of candidate assembly factors are fraught with bottlenecks due to lack of facile assay system to capture assembly defects. Here, we show that bimolecular fluorescence complementation (BiFC) allows detection of assembly defects that are induced by the loss of assembly factors. The fusion of N and C-terminal fragments of Venus fluorescent protein to the ribosomal proteins uS13 and uL5, respectively, in Escherichia coli facilitated the incorporation of the tagged uS13 and uL5 onto the respective ribosomal subunits. When the ribosomal subunits associated to form the 70S particle, the complementary fragments of Venus were br

SUBMITTER: Sharma H 

PROVIDER: S-EPMC5014008 | biostudies-literature | 2016 Sep

REPOSITORIES: biostudies-literature