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Regulation of DNA synthesis at the first cell cycle in the sea urchin in vivo.
- Source :
-
Methods in molecular biology (Clifton, N.J.) [Methods Mol Biol] 2014; Vol. 1128, pp. 223-36. - Publication Year :
- 2014
-
Abstract
- Using fluorescent and non-fluorescent recombinant proteins has proved to be a very successful technique for following postfertilization events, in both male and female pronuclei during the first cell cycle of sea urchin in vivo. Proteins and dyes are introduced by microinjection into the unfertilized egg, and their function can be monitored by fluorescence or confocal/two-photon (2P) and transmitted light microscopy after insemination. Here, we describe expression and purification of GFP/RFP-tagged proteins involved in regulation of DNA replication. We also explain the techniques used to introduce recombinant proteins and fluorescent tubulin into sea urchin eggs and embryos.
- Subjects :
- Animals
Cell Cycle
Cell Nucleus metabolism
Escherichia coli
Female
Fertilization
Fluorescent Dyes chemistry
Fluorescent Dyes isolation & purification
Green Fluorescent Proteins biosynthesis
Green Fluorescent Proteins chemistry
Green Fluorescent Proteins isolation & purification
Male
Microinjections
Microscopy, Confocal
Microscopy, Fluorescence
Ovum cytology
Proliferating Cell Nuclear Antigen biosynthesis
Proliferating Cell Nuclear Antigen chemistry
Proliferating Cell Nuclear Antigen isolation & purification
Sea Urchins physiology
Staining and Labeling
Tubulin chemistry
DNA Replication
Sea Urchins cytology
Subjects
Details
- Language :
- English
- ISSN :
- 1940-6029
- Volume :
- 1128
- Database :
- MEDLINE
- Journal :
- Methods in molecular biology (Clifton, N.J.)
- Publication Type :
- Academic Journal
- Accession number :
- 24567218
- Full Text :
- https://doi.org/10.1007/978-1-62703-974-1_15