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9号染色体辐射杂种的构建与鉴定及其在染色体上定位黏粒探针中的应用。

Construction and characterization of radiation hybrids for chromosome 9, and their use in mapping cosmid probes on the chromosome.

作者信息

Jackson C L, Britt D E, Graw S L, Potts A, Santoro K, Buckler A J, Housman D E, Mark H F

机构信息

Department of Pathology, Rhode Island Hospital, Providence 02903.

出版信息

Somat Cell Mol Genet. 1992 May;18(3):285-301. doi: 10.1007/BF01233864.

Abstract

Radiation hybrids were produced from a monochromosomal microcell hybrid (PK87-9) which contains only human chromosome 9 with an inserted marker on 9p. Doses of radiation ranging from 1000 to 8000 rads were used to produce a series of hybrids with different size fragments of human chromosome 9. The inserted dominant selectable marker was used to select for hybrids that preferentially maintain fragments of 9p. A panel of 53 radiation hybrids were characterized for 17 chromosome 9 markers. In addition, 17 hybrids were analyzed by fluorescent in situ hybridization (FISH). Hybrids were produced with breaks on both 9p and 9q, many of which appear to contain a single fragment of human chromosome 9. These hybrid cell lines were used to regionally localize 31 cosmids isolated from a chromosome 9 cosmid library. Six cosmids were mapped to intervals on 9p, six cosmids mapped to the centromeric region of the chromosome, and 19 mapped to 9q.

摘要

辐射杂种细胞是由一个单染色体微细胞杂种(PK87-9)产生的,该杂种仅包含人类9号染色体,在9号染色体短臂上插入了一个标记。使用1000至8000拉德的辐射剂量来产生一系列具有不同大小人类9号染色体片段的杂种细胞。插入的显性选择标记用于筛选优先保留9号染色体短臂片段的杂种细胞。对一组53个辐射杂种细胞进行了17个9号染色体标记的鉴定。此外,通过荧光原位杂交(FISH)分析了17个杂种细胞。产生的杂种细胞在9号染色体短臂和长臂上均有断裂,其中许多似乎只包含人类9号染色体的一个片段。这些杂种细胞系用于对从9号染色体黏粒文库中分离出的31个黏粒进行区域定位。6个黏粒定位到9号染色体短臂的区间,6个黏粒定位到染色体的着丝粒区域,19个黏粒定位到9号染色体长臂。

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