Laboratory Methods in Enzymology: Protein Part B

Small-scale expression of proteins in E. coliProtein expression-YeastRecombinant protein expression in baculovirus-infected insect cellsSingle cell cloning of a stable mammalian cell lineIn vitro synthesis of proteins in bacterial extractsPreparation of a Saccharomyces cerevisiae cell-free extract for in vitro translationYeast two-hybrid screenUV cross-linking of interacting RNA and protein in cultured cellsAnalysis of RNA-protein interactions by cell mixingGeneral protein-protein cross-linkingChromatin immunoprecipitation and multiplex sequencing (ChIP-Seq) to identify global transcription factor binding sites in the nematode Caenorhabditis elegansPAR-CLIP (photoactivatable ribonucleoside-enhanced crosslinking and immunoprecipitation): A step-by-step protocol to the transcriptome-wide identification of binding sites of RNA-binding proteinsDetermining the RNA specificity and targets of RNA-binding proteins using a three-hybrid systemDissecting a known RNA-protein interaction using a yeast three-hybrid systemIdentifying proteins that bind a known RNA sequence using the yeast three-hybrid system