Large scale isolation of nuclei from oocytes of Xenopus laevis.
We describe an improvement on the procedure of Scalenghe et al. (F. Scalenghe, M. Buscaglia, C. Steinheil, and M. Crippa (1978) Chromosoma 66, 299-308) for the large scale isolation of nuclei from Xenopus laevis oocytes. The nuclear extract obtained was tested for its ability to transcribe a cloned Xenopus 5 S RNA gene and for the presence of nuclear factors interacting with a X. laevis ribosomal protein gene promoter. Efficiency of accurate transcription and of factor binding is comparable with that of an extract prepared from manually isolated nuclei.
Ruberti
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Dipartimento di Genetica e Biologia Molecolare, Università di Roma La Sapienza, Italy.
Beccari
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Bianchi
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Carnevali
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F
D000818:Animals; D002458:Cell Fractionation / Q000379:methods*; D002467:Cell Nucleus / Q000378:metabolism*; D015342:DNA Probes; D004268:DNA-Binding Proteins / Q000032:analysis; D005260:Female; D066298:In Vitro Techniques; D009865:Oocytes / Q000378:metabolism / Q000648:ultrastructure*; D010957:Plasmids; D011401:Promoter Regions, Genetic; D012341:RNA, Ribosomal, 5S / Q000235:genetics; D014158:Transcription, Genetic; D014982:Xenopus laevis
D015342:DNA Probes; D004268:DNA-Binding Proteins; D012341:RNA, Ribosomal, 5S