Production of a lampbrush chromosome

C - Chemistry – Metallurgy – 12 – Q

Patent

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C12Q 1/68 (2006.01)

Patent

CA 2277804

A process for inducing a transcriptionally active chromosome (i.e., a "lampbrush" chromosome) from condensed chromatin or nucleus is disclosed. The condensed chromosome is contacted with the contents of a germinal vesicle. Preferably, the nuclear envelope is disrupted or removed. Moreover, a heterologous system of chromosome and germinal vesicle derived from organisms of different species is preferred because it permits analysis of organisms that do not have lampbrush chromosomes or cannot be manipulated by other techniques. Such lampbrush chromosomes can be attached to a substrate, and then analyzed by a variety of molecular and cytological techniques such as, for example, antibody detection of chromosomal protein, autoradiography, electron and light microscopy, histochemistry, image analysis, immunofluorescence, in situ hybridization of nucleic acids, morphology, and the like.

L'invention concerne un procédé qui permet d'induire un chromosome à activité transcriptionnelle (c'est-à-dire un chromosome plumeux) à partir de noyau ou de chromatine sous forme condensée. On met en contact le chromosome condensé avec le contenu d'une vésicule germinative. De préférence, l'enveloppe du noyau est rompue ou éliminée. De plus, on utilise de préférence un système hétérologue de chromosome et de vésicule germinative dérivé d'organismes d'espèces différentes, car cela permet d'analyser des organismes dépourvus de chromosomes plumeux ou ne pouvant être manipulés via d'autres techniques. Lesdits chromosomes peuvent être fixés à un substrat, puis analysés par le biais d'un éventail de techniques moléculaires et cytologiques, à savoir entre autres : détection d'anticorps sur protéine chromosomique, autoradiographie, microscopie électronique ou photonique, histochimie, analyse d'image, immunofluorescence, hybridation in situ d'acides nucléiques, morphologie, etc.

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