Exogenous protein expression in Xenopus oocytes: Basic procedures
Bossi, E.; Fabbrini, M.S.; Ceriotti, A. (2007). Exogenous protein expression in Xenopus oocytes: Basic procedures, in: Grandi, G. (Ed.) In vitro transcription and translation protocols. Methods in Molecular Biology, 375: pp. 107-131. https://dx.doi.org/10.1007/978-1-59745-388-2_6
In: Grandi, G. (Ed.) (2007). In vitro transcription and translation protocols. Second Edition. Methods in Molecular Biology, 375. Second Edition. Humana Press: Totowa. ISBN 978-1-58829-558-3. , more
In: Methods in Molecular Biology. Humana Press. ISSN 1064-3745, more
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| Keywords |
Cells > Sexual cells > Eggs > Oocytes
Xenopus laevis
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| Author keywords |
mRNA; translation; microinjection |
| Authors | | Top |
- Bossi, E.
- Fabbrini, M.S.
- Ceriotti, A.
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| Abstract |
The oocytes of the South African clawed frog Xenopus laevis have been widely used as a reliable system for the expression and characterization of different types of proteins, including ion channels and membrane receptors. The large size and resilience of these oocytes make them easy to handle and to microinject with different molecules such as natural mRNAs, cRNAs, and antibodies. A variety of methods can then be used to monitor the expression of the proteins encoded by the microinjected mRNA/cRNA, and to perform a functional characterization of the heterologous polypeptides. In this chapter, after describing the equipment required to maintain X. laevis in the laboratory and to set up a microinjection system, we provide detailed procedures for oocyte isolation, micropipet and cRNA preparation, and oocyte microinjection. A method for the labeling of oocyte-synthesized proteins and for the immunological detection of the heterologous polypeptides is also described. |
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