A 350-kb cosmid contig in 3p14.2 that crosses the t(3;8) hereditary renal cell carcinoma translocation breakpoint and 17 aphidicolin-induced FRA3B breakpoints.
A 350-kb cosmid contig in 3p14.2 that crosses the t(3;8) hereditary renal cell carcinoma translocation breakpoint and 17 aphidicolin-induced FRA3B breakpoints.
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3p14.2 中的 350 kb 粘粒重叠群跨越 t(3;8) 遗传性肾细胞癌易位断点和 17 个阿非迪霉素诱导的 FRA3B 断点。
DOI：
10.1006/geno.1996.0326
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发表时间：
1996
期刊：
影响因子：
--
通讯作者：
Smith,DI
中科院分区：
文献类型：
--
作者：
Paradee,W;Wilke,CM;Wang,L;Shridhar,R;Mullins,CM;Hoge,A;Glover,TW;Smith,DI
人染色体3p14.2区的组成型脆性位点FRA 3B被认为是人类基因组中最活跃的脆性位点。FRA 3B在细胞学上与遗传性肾细胞癌（hRCC）易位t（3;8）（p14.2;q24.13）中观察到的3号染色体断裂点无法区分。先前的工作表明，一个1330 kb的YAC克隆，YC 850 A6，跨越t（3;8）易位和FRA 3B，也包括FRA 3B相关的断点诱导仓鼠-人杂交。该YAC用于构建多命中粘粒文库。该文库的筛选导致从t（3;8）易位断裂点向远端延伸的350-kb粘粒重叠群。17个aphidicolin诱导的3p14.2断裂点来自仓鼠-人的杂种在这个粘粒重叠群中进行了定位。发现这些断裂点定位为两个不同的簇，相隔200 kb，位于FRA 3B内频繁断裂区域的两侧，如使用来自重叠群的互补链进行FISH分析所定义。断裂点簇的最近端位于hRCC t（3;8）断裂点远侧约100 kb处。这些断裂点的分布，以及通过FISH分析绘制的频繁染色体断裂区域，进一步证实了FRA 3B的位置，并有助于确定其脆性的发挥程度。这些数据表明，FRA 3B在3p14.2内包含数百个DNA序列的碱基序列。从YAC YC 850 A6构建的350-kb重叠群和粘粒文库对于进一步表征FRA 3B周围区域和确定FRA 3B脆性的分子基础的实验将是必不可少的。
The constitutive fragile site at human chromosomal band 3p14.2, FRA3B, has been described as the most active common fragile site in the human genome. FRA3B is cytologically indistinguishable from the chromosome 3 breakpoint observed in the hereditary renal cell carcinoma (hRCC) translocation t(3;8) (p14.2;q24.13). Previous work demonstrated that a 1330-kb YAC clone, YC850A6, spans both the t(3;8) translocation and FRA3B and also encompasses FRA3B-associated breakpoints induced in hamster–human hybrids. This YAC was used to construct a multi-hit cosmid library. Screening of this library resulted in a 350-kb cosmid contig that extends distally from the t(3;8) translocation breakpoint. Seventeen aphidicolin-induced 3p14.2 breakpoints derived from hamster–human hybrids were mapped within this cosmid contig. These breakpoints were found to localize as two distinct clusters, separated by 200 kb, which lie on either side of a region of frequent breakage within FRA3B as defined by FISH analysis using cosmids from the contigs. The most proximal of the breakpoint clusters lies approximately 100 kb distal to the hRCC t(3;8) breakpoint. The distribution of these breakpoints, together with the region of frequent chromosomal breakage mapped by FISH analysis, further confirms the position of FRA3B and helps to define the extent over which its fragility is exerted. These data indicate that FRA3B comprises several hundred kilobases of DNA sequence within 3p14.2. The 350-kb contig and the cosmid library constructed from YAC YC850A6 will be essential for further characterization of the region surrounding FRA3B and in experiments to determine the molecular basis of the fragility of FRA3B.