Abstract
Fluorescently tagged proteins have become a crucial weapon in the armory of a successful cell biology laboratory. This chapter describes how to produce cRNA coding for a fluorescently tagged protein of choice, such that it is suitable for microinjection and subsequent expression studies in live oocytes.
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References
Prasher DC (1995) Using GFP to see the light. Trends Genet 11:320–323
Shaner NC, Steinbach PA, Tsien RY (2005) A guide to choosing fluorescent proteins. Nat Methods 2:905–909
Day RN, Davidson MW (2009) The fluorescent protein pallette: tools for cellular imaging. Chem Soc Rev 38:2887–2921
Whitaker M (1996) Control of meiotic arrest. Rev Reprod 1:127–135
Levasseur M, McDougall A (2000) Sperm-induced calcium oscillations at fertilisation in ascidians are controlled by cyclin B1-dependent kinase activity. Development 127:631–641
Herbert M, Levasseur M, Homer H, Yallop K, Murdoch A, McDougall A (2003) Homologue disjunction in mouse oocytes requires proteolysis of securin and cyclin B1. Nat Cell Biol 5:1023–1025
Lemaire P, Garret N, Gurdon JB (1995) Expression cloning of Siamois, a Xenopus homeobox gene expressed in dorsal-vegetal cells of blastulae and able to induce a complete secondary axis. Cell 81:85–94
Kreig PA, Melton DA (1984) Functional messengers are produced by SP6 in vitro transcription of cloned cDNAs. Nucleic Acids Res 12:7057–7070
Homer HA, McDougall A, Levasseur M, Yallop K, Murdoch AP, Herbert M (2005) Mad2 prevents aneuploidy and premature proteolysis of cyclin B and securin during meiosis I in mouse oocytes. Genes Dev 19:202–207
Zernicka-Goetz M, Pines J, MacLean Hunter S, Dixon JPC, Siemering KR, Hasselhof J, Evans MJ (1997) Following cell fate in the living mouse embryo. Development 124:1133–1137
Mullis KB, Faloona F (1987) Specific synthesis of DNA in vitro via a polymerase catalyzed chain reaction. Methods Enzymol 155:335–350
Wentzell LM, Nobbs TJ, Halford SE (1995) The SfiI restriction endonuclease makes a four-strand DNA break at two copies of its recognition sequence. J Mol Biol 248:581–595
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Levasseur, M. (2013). Making cRNA for Microinjection and Expression of Fluorescently Tagged Proteins for Live-Cell Imaging in Oocytes. In: Homer, H. (eds) Mammalian Oocyte Regulation. Methods in Molecular Biology, vol 957. Humana Press, Totowa, NJ. https://doi.org/10.1007/978-1-62703-191-2_8
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DOI: https://doi.org/10.1007/978-1-62703-191-2_8
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Publisher Name: Humana Press, Totowa, NJ
Print ISBN: 978-1-62703-190-5
Online ISBN: 978-1-62703-191-2
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