In vivo selection of functional variations in essential sites of ribosomal RNA.
The technique of "in vivo selection of functional ribosomes" is a genetic approach to dissecting the link between the structure and function of critical sites of rRNA. This method proceeds through selection of functional variants among cells that express ribosomes from a pool of rRNA-containing randomized sites. The selection of bacterial clones with functional ribosomes is based on the use of a plasmid carrying a rRNA operon in which a site of interest has been randomized and a point mutation conferring an antibiotic resistance has been introduced. Cells expressing functional ribosomes are then selected on medium containing the antibiotic. With this approach one can isolate at once all the possible variations at a given rRNA site that are able to sustain normal ribosome function. The identification of covariations in between several nucleotides that maintain wild-type ribosome activity can thus help demonstrate the function of specific interactions in rRNA.
Moine
H
H
UPR 9002 du CNRS, 67084 Strasbourg, France. h.moine@ibmc.u-strasbg.fr
Ehresmann
B
B
Ehresmann
C
C
D001665:Binding Sites; D004926:Escherichia coli / Q000378:metabolism; D005821:Genetic Techniques; D008957:Models, Genetic; D010802:Phylogeny; D010957:Plasmids / Q000378:metabolism; D011485:Protein Binding; D012335:RNA, Ribosomal / Q000737:chemistry* / Q000648:ultrastructure*; D012270:Ribosomes / Q000737:chemistry*
D012335:RNA, Ribosomal