Oocyte expression with injection of purified T7 RNA polymerase.
The Xenopus oocyte is a widely used system for protein expression. Investigators have had the choice between two different techniques: injection into the cytoplasm of in vitro transcribed complementary RNA (cRNA) or injection into the nucleus of complementary DNA (cDNA). We report on a third expression technique that is based on the combined injection of cDNA and purified T7 RNA polymerase directly into the cytoplasm of oocytes.
Altafaj
Xavier
X
Département Réponse et Dynamique Cellulaire, Inserm U607, Grenoble, France.
Joux
Nathalie
N
Ronjat
Michel
M
De Waard
Michel
M
D000818:Animals; D017123:Bacteriophage T7 / Q000201:enzymology; D012321:DNA-Directed RNA Polymerases / Q000737:chemistry / Q000302:isolation & purification; D005260:Female; D008845:Microinjections / Q000295:instrumentation / Q000379:methods; D036103:Nanotechnology / Q000295:instrumentation / Q000379:methods; D009865:Oocytes / Q000378:metabolism*; D014764:Viral Proteins / Q000737:chemistry / Q000302:isolation & purification; D014982:Xenopus laevis
D014764:Viral Proteins; C062210:bacteriophage T7 RNA polymerase; D012321:DNA-Directed RNA Polymerases