Laboratory Methods in Enzymology: Protein Part B

Small-scale expression of proteins in E. coli Protein expression-Yeast Recombinant protein expression in baculovirus-infected insect cells Single cell cloning of a stable mammalian cell line In vitro synthesis of proteins in bacterial extracts Preparation of a Saccharomyces cerevisiae cell-free extract for in vitro translation Yeast two-hybrid screen UV cross-linking of interacting RNA and protein in cultured cells Analysis of RNA–protein interactions by cell mixing General protein–protein cross-linking Chromatin immunoprecipitation and multiplex sequencing (ChIP-Seq) to identify global transcription factor binding sites in the nematode Caenorhabditis elegans PAR-CLIP (photoactivatable ribonucleoside-enhanced crosslinking and immunoprecipitation): A step-by-step protocol to the transcriptome-wide identification of binding sites of RNA-binding proteins Determining the RNA specificity and targets of RNA-binding proteins using a three-hybrid system Dissecting a known RNA–protein interaction using a yeast three-hybrid system Identifying proteins that bind a known RNA sequence using the yeast three-hybrid system

Biochemists, biophysicists, molecular biologists, analytical chemists, and physiologists

• Indispensable tool for the researcher
• Carefully written and edited by experts to contain step-by-step protocols
• In this volume we have brought together a number of core protocols concentrating on protein