Single-embryo RT-PCR assay to study gene expression dynamics during embryogenesis inArabidopsis thaliana
Purchase on Springer.com
$39.95 / €34.95 / £29.95*
Rent the article at a discountRent now
* Final gross prices may vary according to local VAT.
We have developed a single-embryo RT-PCR protocol for studying gene expression during plant embryogenesis. Four genes,glyceraldhyde-3-phosphate dehydrogenase (GAPC), shoot-meristemless (STM), monopteros (MP), andshaggy-like kinase etha (ASKη), fromArabidopsis thaliana were used to test the sensitivity and reliability of this method by analyzing the differential signal intensities of their RT-PCR products. The method could detect genes expressed during embryogenesis at a single-embryo level and, therefore, can be used to identify phenotypes. When in vitro, embryogenesis also is used to control the time course of zygote development exactly. The single-embryo RT-PCR protocol becomes a powerful method to survey the dynamics of specific gene expression.
- Adjaye J, Bolton V, and Monk M (1999) Development expression of specific genes detected in high-quality cDNA library from single human preimplantation embryos. Gene 237: 373–83. CrossRef
- Dornelas MC and Kreis M (1997) Three New cDNAs Related to SGG/GSK-3 (SHAGGY/Glycogen Synthase Kinase-3) fromArabidopsis thaliana. Plant Physiol 113: 306–306.
- Faure JE, Digonnet C, and Dumas C (1994) An in vitro system for adhesion and fusion of maize gametes. Science 263: 1598–160. CrossRef
- Hardtke CS and Berleth T (1998) The Arabidopsis gene MONOPTEROS encodes a transcription factor mediating embryo axis formation and vascular development. EMBO J 17(5): 1405–1411. CrossRef
- Jacqueline KP and Lipski J (2000) Single-cell RT-PCR as a tool to study gene expression in central and peripheral autonomic neurons. Autonomic Neuroscience: Basic and Clinical 86: 1–12. CrossRef
- Johansen FF, Lambolez B, Audinat E, Bochet P, and Rossier J (1995) Single cell RT-PCR proceeds without the risk of genomic DNA amplification. Neurochem Int 26: 239–243. CrossRef
- Karrer EE, Lincoln JE, Hogenhout S, Bennett AB, Bostock RM, MarTineau B, Lucas WJ, Gilchrist DG, and Alexander D (1995) In situ isolation of mRNA from individual plant cells: creation of cell-specific cDNA library. Proc Natl Acad Sci 92: 3814–3818. CrossRef
- Kawasaki E, Hattori N, Miyamoto E, Yamashita T, and Inagaki C (1999) Single-cell RT-PCR demonstrates expression of voltage-dependent chloride channels (CIC-1, CIC-2 and CIC-3) in outer hair cells of rat cochlea. Brain Research 838: 166–170. CrossRef
- Kranz E, Bautor J, and Lorz H (1991) In vitro fertilization with isolated, single gametes by electrofusion. Sex Plant Reprod 4: 12–16.
- Long JA and Barton MK (1996) A member of the KNOTTED class of homeodomain proteins encoded by the STM gene ofArabidopsis. Nature 379(6560): 66–69. CrossRef
- Preuss D (1999) Chromatin silencing andArabidopsis development: a role for Polycomb protein. Plant Cell 11: 765–767.
- Richert J, Kranz E, Lörz H, and Dresselhaus T (1996) A transcriptase-polymerase chain reaction assay for gene expression studies at the single cell level. Plant Science 114: 93–99. CrossRef
- Shih MC and Goodman HM (1991) Cloning and chromosomal mapping of Nuclear genes encoding chloroplast and cytosolic glyceraldehyde-3-phosphate dehydrogenase fromArabidopsis thaliana. Gene 104: 133–138. CrossRef
- Sun M-X, Moscatelli A, Yang H-Y, and Cresti M (2000) In vitro double fertilization inNicotiana tabacum (L.): fusion behavior and gamete interaction traced by video-enhanced microscopy. Sex Plant Reprod 12: 267–275. CrossRef
- Single-embryo RT-PCR assay to study gene expression dynamics during embryogenesis inArabidopsis thaliana
Plant Molecular Biology Reporter
Volume 20, Issue 1 , pp 19-26
- Cover Date
- Print ISSN
- Online ISSN
- Additional Links
- Arabidopsis thaliana
- gene expression
- single embryo
- Industry Sectors