A novel emulsion mixture for in vitro compartmentalization of transcription and translation in the rabbit reticulocyte system.
Emulsion formulations used for in vitro compartmentalization (IVC) methods were found to be incompatible with protein expression in the rabbit reticulocyte (RRL) system, causing rapid discoloration and translation shutdown. Here we identify possible causes and describe a novel water-in-oil emulsion which abolished discoloration and allowed high-level in-emulsion expression of active luciferase and human telomerase using the RRL. This novel emulsion greatly expands the range of potential protein targets for IVC.
MRC Laboratory of Molecular Biology, Hills Road, Cambridge CB2 2QH, UK.
Farid J
FJ
Ghadessy
Philipp
P
Holliger
D000818:Animals; D002474:Cell-Free System; D045703:Emulsifying Agents / Q000737:chemistry; D004655:Emulsions / Q000737:chemistry*; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D008899:Mineral Oil / Q000737:chemistry; D014176:Protein Biosynthesis; D011817:Rabbits; D012156:Reticulocytes / Q000201:enzymology*; D012996:Solutions; D019098:Telomerase / Q000235:genetics / Q000378:metabolism; D014158:Transcription, Genetic; D014867:Water / Q000737:chemistry
D045703:Emulsifying Agents; D004655:Emulsions; D012996:Solutions; D014867:Water; D008899:Mineral Oil; D008156:Luciferases; D019098:Telomerase