Oocyte expression with injection of purified T7 RNA polymerase

Methods Mol Biol. 2006:322:55-67. doi: 10.1007/978-1-59745-000-3_5.

Abstract

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.

Publication types

  • Review

MeSH terms

  • Animals
  • Bacteriophage T7 / enzymology
  • DNA-Directed RNA Polymerases* / chemistry
  • DNA-Directed RNA Polymerases* / isolation & purification
  • Female
  • Microinjections* / instrumentation
  • Microinjections* / methods
  • Nanotechnology / instrumentation
  • Nanotechnology / methods
  • Oocytes / metabolism*
  • Viral Proteins* / chemistry
  • Viral Proteins* / isolation & purification
  • Xenopus laevis

Substances

  • Viral Proteins
  • bacteriophage T7 RNA polymerase
  • DNA-Directed RNA Polymerases