Echinococcus granulosus: preparation of protein extracts from protoscolex nuclei for mobility-shift assays.
Nuclei from Echinococcus granulosus protoscolices were isolated from infected sheep. Protein extracts were prepared for analysis of DNA-protein interactions involving specific transcriptional regulatory factors. Gel mobility-shift assays were done using a heterologous probe containing binding sites for widespread transcription factors. A fragment of the promoter of GATA-1 transcription factor from the chicken was selected. When nuclear extracts from E. granulosus protoscolices were assayed a specific band shift was observed. The methodologies developed in this study could provide an important contribution for the characterization of the DNA-protein interactions involved in transcriptional regulation within the context of recent developments in the molecular biology of this parasite.
Departamento de Bioquímica, Facultad de Ciencias, Montevideo, Uruguay. bgarat@genetica.edu.uy
B
B
Garat
P
P
Esperón
M
M
Picón
R
R
Ehrlich
D000818:Animals; D001345:Autoradiography; D002458:Cell Fractionation; D002467:Cell Nucleus / Q000737:chemistry; D002645:Chickens; D004247:DNA / Q000378:metabolism; D004268:DNA-Binding Proteins / Q000235:genetics / Q000302:isolation & purification* / Q000378:metabolism; D004446:Echinococcus / Q000737:chemistry*; D004591:Electrophoresis, Polyacrylamide Gel; D050981:Erythroid-Specific DNA-Binding Factors; D015801:Helminth Proteins / Q000302:isolation & purification* / Q000378:metabolism; D009687:Nuclear Proteins / Q000302:isolation & purification* / Q000378:metabolism; D011401:Promoter Regions, Genetic; D012756:Sheep; D014157:Transcription Factors / Q000235:genetics
D004268:DNA-Binding Proteins; D050981:Erythroid-Specific DNA-Binding Factors; D015801:Helminth Proteins; D009687:Nuclear Proteins; D014157:Transcription Factors; D004247:DNA