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人类3号染色体短臂上80个黏粒克隆的分离与荧光原位杂交定位

Isolation and FISH mapping of 80 cosmid clones on the short arm of human chromosome 3.

作者信息

Haas M, Aburatani H, Stanton V P, Bhatt M, Housman D, Ward D C

机构信息

Department of Genetics, Yale University School of Medicine, New Haven, Connecticut 06510.

出版信息

Genomics. 1993 Apr;16(1):90-6. doi: 10.1006/geno.1993.1145.

Abstract

We have isolated and mapped by fluorescence in situ hybridization 80 new cosmids on the short arm of chromosome 3. These markers were isolated from a radiation-reduced hybrid, DM1, made from a cell line that was monochromosomal for human chromosome 3. Selected cosmids were used in double-label cohybridization experiments in which polymerase chain reaction products, generated by an Alu oligonucleotide primer from genomic DNA, were used for chromosome banding. Fifty-six of the cosmid probes map between 3p14.3 and 3p22 while 24 other probes cluster around bands 3p23-3p25. Three probes that appeared to map close to the chromosome 3 region bearing a t(3,8)p14.2; q24.1 translocation associated with renal cell carcinoma were analyzed by interphase mapping techniques and hybridized to metaphase spreads from the translocation cell line. These 80 probes will be useful in the elucidation of genetic alterations associated with diseases such as small cell lung carcinoma, renal cell carcinoma, and von Hippel-Lindau disease.

摘要

我们通过荧光原位杂交技术在3号染色体短臂上分离并定位了80个新的黏粒。这些标记是从一个辐射减少的杂种细胞系DM1中分离出来的,该细胞系是人类3号染色体的单染色体细胞系。选择的黏粒用于双标记共杂交实验,其中由来自基因组DNA的Alu寡核苷酸引物产生的聚合酶链反应产物用于染色体显带。56个黏粒探针定位在3p14.3和3p22之间,而其他24个探针聚集在3p23 - 3p25带周围。通过间期定位技术分析了三个似乎定位在与肾细胞癌相关的t(3,8)p14.2; q24.1易位的3号染色体区域附近的探针,并将其与来自易位细胞系的中期染色体铺展杂交。这80个探针将有助于阐明与小细胞肺癌、肾细胞癌和冯·希佩尔-林道病等疾病相关的基因改变。

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