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牛的双变量流式核型分析以及利用PCR生成的探针进行染色体涂染的峰识别。

The bovine bivariate flow karyotype and peak identification by chromosome painting with PCR-generated probes.

作者信息

Schmitz A, Oustry A, Chaput B, Bahri-Darwich I, Yerle M, Millan D, Frelat G, Cribiu E P

机构信息

Centre d'études nucléaires de Fontenay aux roses (CEA), Direction des Sciences du vivant, Laboratoire de Cytométrie, France.

出版信息

Mamm Genome. 1995 Jun;6(6):415-20. doi: 10.1007/BF00355643.

Abstract

A bovine bivariate flow karyotype has been established from a primary fibroblast cell culture carrying a 4;10 Robertsonian translocation. From 27 to 36 populations could be resolved by flow cytometry although the anticipated number was 31. Separation of chromosomal pairs into two populations explains this high resolution and confirms the high level of heteromorphism previously observed. We used a PARM-PCR (Priming Authorizing Random Mismatches) procedure for the production of paint probes from flow-sorted chromosome fractions. These probes were used for chromosome identification by fluorescence in situ hybridization (FISH) on R-banded metaphase spreads. We present the localization of all the bovine chromosome types on the flow karyotype. Twenty-two chromosome types including the translocated chromosome were sorted as pure fractions.

摘要

已从携带4;10罗伯逊易位的原代成纤维细胞培养物中建立了牛双变量流式核型。尽管预期数量为31,但通过流式细胞术可分辨出27至36个群体。染色体对分离为两个群体解释了这种高分辨率,并证实了先前观察到的高度异质性。我们使用PARM-PCR(引物授权随机错配)程序从流式分选的染色体组分中制备涂染探针。这些探针用于通过R带中期铺展上的荧光原位杂交(FISH)进行染色体鉴定。我们展示了所有牛染色体类型在流式核型上的定位。包括易位染色体在内的22种染色体类型被分选成纯组分。

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