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用于流式细胞术和分选的异常猪染色体的鉴定及细胞遗传学分析。

Identification and cytogenetic analysis of an abnormal pig chromosome for flow cytometry and sorting.

作者信息

Hausmann M, Popescu C P, Boscher J, Kerboeuf D, Dölle J, Cremer C

机构信息

Laboratoire de Cytogénétique, INRA, Jouy-en-Josas, France.

出版信息

Z Naturforsch C J Biosci. 1993 Jul-Aug;48(7-8):645-53. doi: 10.1515/znc-1993-7-819.

Abstract

For cytogenetics of pig (Sus scrofa domestica) and the influence of chromosome aberrations on pig production, high interest exists in flow sorted chromosomes for gene mapping, to establish DNA-libraries, or to produce DNA-probes. Flow karyotyping and sorting as well as slit scan flow analysis of metaphase chromosomes of an abnormal cell type carrying a translocation marker chromosome 6/15 are described. Flow sorting of the largest chromosomes of these cells was performed. After sorting the chromosomes still had a well preserved morphology and were identified microscopically by G-banding. The quality of the band pattern of the sorted chromosomes was compatible to that of isolated chromosomes not subjected to flow cytometry. The sorted fraction showed an enrichment of chromosome 6/15 and chromosome 1 which have quantitatively about the same integrated fluorescence intensity. Slit scan flow analysis was performed to discriminate these two chromosomes. Metacentric and submetacentric chromosomes were analyzed according to their bimodal slit scan profiles. Profiles of the largest chromosomes were distinguished by their different centromeric indices. Two groups were interpreted as the normal chromosome 1 and the translocation chromosome 6/15.

摘要

关于猪(家猪)的细胞遗传学以及染色体畸变对猪生产的影响,人们对用于基因定位、建立DNA文库或制备DNA探针的流式分选染色体有着浓厚的兴趣。本文描述了对携带易位标记染色体6/15的异常细胞类型的中期染色体进行流式核型分析和分选以及狭缝扫描流式分析的过程。对这些细胞中最大的染色体进行了流式分选。分选后的染色体仍具有保存良好的形态,并通过G带显微镜鉴定。分选染色体的带型质量与未进行流式细胞术处理的分离染色体的带型质量相当。分选部分显示出6/15染色体和1号染色体的富集,它们在定量上具有大致相同的整合荧光强度。进行狭缝扫描流式分析以区分这两条染色体。根据其双峰狭缝扫描图谱分析了中着丝粒和亚中着丝粒染色体。最大染色体的图谱通过其不同的着丝粒指数来区分。两组分别被解释为正常的1号染色体和易位染色体6/15。

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