Paternal gene expression in developing hybrid embryos of Xenopus laevis and Xenopus borealis

Development ◽  
1980 ◽  
Vol 60 (1) ◽  
pp. 359-372
Author(s):  
H. R. Woodland ◽  
J. E. M. Ballantine
Keyword(s):  
Gene Expression ◽  
Xenopus Laevis ◽  
Gel Electrophoresis ◽  
2D Gel Electrophoresis ◽  
Gastrula Stage ◽  
Maternal Mrna ◽  
Specific Proteins ◽  
2D Gel ◽  
Androgenetic Haploid ◽  
Xenopus Borealis

We have studied protein synthesis in the viable hybrid Xenopus laevis (♀) × Xenopus borealis (♂) using 2D gel electrophoresis. Fourteen borealis-specific proteins were studied. Two of these proteins appeared by the gastrula stage, five in the gastrula and the rest later. Where homologous laevis proteins were tentatively identified, androgenetic haploid hybrids were used to study whether the protein was encoded by stored maternal mRNA, and how long this mRNA persisted. The two proteins appearing in blastulae were probably initially coded by stored maternal mRNA. This was not detectable by the tailbud-tadpole stage, and presumably had been destroyed.

Protein synthesis and messenger RNA levels along the animal–vegetal axis during early Xenopus development

Development ◽  
1986 ◽  
Vol 95 (1) ◽  
pp. 15-35
Author(s):  
Rosamund C. Smith
Keyword(s):  
Protein Synthesis ◽  
Gel Electrophoresis ◽  
Messenger Rna ◽  
Xenopus Oocytes ◽  
Early Embryo ◽  
2D Gel Electrophoresis ◽  
Gastrula Stage ◽  
Cleavage Stage ◽  
2D Gel

The patterns of proteins synthesized in animal and vegetal regions of Xenopus oocytes, eggs and embryos were examined by 2D gel electrophoresis. In oocytes and eggs, the only proteins synthesized asymmetrically along the animal-vegetal axis were a small number of proteins synthesized predominantly in the vegetal hemisphere. At the cleavage stage there were a total of four proteins synthesized unevenly in animal and vegetal regions: three synthesized predominantly in the vegetal hemisphere and one synthesized predominantly in the animal hemisphere. By the gastrula stage, when maternal messages have largely been replaced by embryonic transcripts, the number of differences in proteins synthesized in the animal-derived ectoderm and mesoderm, and the vegetal-derived endoderm started to increase rapidly with time of development with many more animal-characteristic proteins than vegetal-characteristic proteins appearing. Comparison of protein synthesis patterns with those obtained when extracted RNA was translated in vitro and run on 2D gels, showed that the asymmetry in protein synthesis along the animal-vegetal axis in the oocyte and early embryo reflected directly the distribution of their mRNAs along the axis. There was no evidence for localized ‘masked’ abundant messages along the animal-vegetal axis of oocytes and cleavage embryos.

Identification of nuclei associated proteins by 2D-gel electrophoresis and mass spectrometry

2001 ◽  
Vol 109 (1) ◽  
pp. 3-11 ◽  
Author(s):  
Jonas Bergquist ◽  
Johan Gobom ◽  
Anders Blomberg ◽  
Peter Roepstorff ◽  
Rolf Ekman
Keyword(s):  
Mass Spectrometry ◽  
Gel Electrophoresis ◽  
2D Gel Electrophoresis ◽  
Associated Proteins ◽  
2D Gel

scholarly journals Differential methods of cell lysis for protein profiling of Alexandrium sp. using 2D gel electrophoresis

2020 ◽  
Vol 15 (1) ◽  
Author(s):  
Nurul Ashima Hamdan ◽  
Normawaty Mohammad-Nor ◽  
Shafida Abd. Hamid ◽  
Nor Hasniza Md. Zin ◽  
Noraslinda Muhamad Bunnori
Keyword(s):  
Gel Electrophoresis ◽  
Cold Water ◽  
Protein Extraction ◽  
Excellent Result ◽  
Axenic Culture ◽  
Cell Lysis ◽  
Protein Profiling ◽  
2D Gel Electrophoresis ◽  
Fundamental Study ◽  
2D Gel

Introduction: Protein profiling of harmful algae is an ongoing study where the latest analysis was conducted on A. minutum. It is a fundamental study where the protein expression of targeted species can be used to understand the biochemical pathway of selected proteins. The Malaysia Alexandrium spp. has the potential to cause massive blooming that brings harm to the aquatic ecosystem. Methods: In this experiment three methods of cell lysis were tested against A. leei isolated from Malaysian waters. Results: The Axenic culture of the sample was established in enriched seawater media (ESDK) with 12 hours of light and 12 hours of dark conditions. The sample was extracted during exponential phase (day 18) where the same amount of cells was collected via centrifugation. The same buffer was used for each technique of cell lysis in order to get the best protein profiles in terms of a band or spot intensity and number. The cells of A. leei were lysed using sonication in the ice-cold water bath, sonication with probe and freeze-thawing followed by sonication in iced bath is the best method of protein extraction. Conclusions: Nevertheless, it can be concluded that the qualitative analysis using SDS-PAGE and 2D gel electrophoresis could be best method and the freeze thawing mode of cell lysis produced an excellent result among others as the protein spots produced were precise and less streaking were observed.

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