Evi-1 expression in Xenopus
Section snippets
Molecular cloning of Xenopus Evi1
A full-length cDNA was isolated from a Xenopus oocyte lambda phage library using a murine c-Evi-1 cDNA probe. The xEvi-1 cDNA was 4208 bp, with 235 bp of 5′-noncoding and 808 bp of 3′-noncoding sequence. The predicted amino acid sequence of the single open reading frame was 1051 amino acids in length and encodes a protein with a calculated molecular mass of 119 kD. Comparison of the human, mouse and Xenopus Evi-1 proteins indicates extensive amino acid identity (Fig. 1). Xenopus Evi-1 shares 76.6%
Oocytes and embryos
Ovaries were manually dissected and treated with collagenase type I (2 mg/ml) in modified Barth solution (MBS) for 15 h. Oocytes (from I to VI) were staged according to Dumont (Dumont, 1972). Stage VI oocytes were matured in vitro by treatment with 5 μg/ml progesterone for 17 h. Albino embryos for in situ hybridization were fertilized in vitro and staged according to Nieuwkoop and Faber (1967)).
cDNA cloning
A Xenopus oocyte cDNA library was screened using the first DNA binding domain of murine Evi-1 (700 bp Sph1
Acknowledgements
We thank Doug Melton for the gift of the Xenopus oocyte λ phage library. James Ihle is an Investigator with the HHMI. We thank Drs Joanne Doherty and Clair Kelley for careful review of this manuscript. This works was supported by the American Lebanese Syrian Associated Charities (ALSAC) and by grants from the NIH (5RO1 DK42932).
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2007, GeneCitation Excerpt :EVI1 orthologs also have specific functions in the neuronal development of invertebrates: the Drosophila EVI1 (or more precisely, MDS1/EVI1, see Section 4) ortholog hamlet specified single versus multiple dendrite morphology in the fly's peripheral nervous system (Moore et al., 2002); and the Caenorhabditis elegans EVI1 ortholog egl-43 was required for the migration of the hermaphrodite specific neurons (HSNs), and for proper morphological development of the PHA and PHB sensory neurons (Garriga et al., 1993). The recently reported expression patterns of Evi1 in Xenopus, chicken, and zebrafish demonstrated that many aspects of the developmental regulation of the Evi1 gene have been conserved throughout vertebrate evolution (Mead et al., 2005; Van Campenhout et al., 2006). As in the mouse, temporal and spatial specificity of Evi1 expression in these organisms argued for a role of Evi1 in development, which was experimentally confirmed for the formation of the Xenopus pronephros (Van Campenhout et al., 2006).