The region surrounding the PKD1 gene: a 700-kb P1 contig from a YAC-deficient interval.
The region surrounding the PKD1 gene: a 700-kb P1 contig from a YAC-deficient interval.
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PKD1 基因周围的区域：来自 YAC 缺陷区间的 700 kb P1 重叠群。
DOI：
10.1101/gr.6.6.515
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发表时间：
1996
期刊：
影响因子：
7
通讯作者：
Klinger,KW
中科院分区：
文献类型：
--
作者：
Dackowski,WR;Connors,TD;Bowe,AE;StantonJr,V;Housman,D;Doggett,NA;Landes,GM;Klinger,KW
作为鉴定多囊肾病（PKD1）主要形式的基因的一部分，我们使用网格人类P1文库进行contig组装。感兴趣的区间是16p13.3染色体上一个700 kb的片段，可以通过遗传标记D16S125和D16S84进行物理描绘，染色体特征为富含gc的等质粒，富含CpG岛、基因和alu样重复。我们试图恢复编码16号染色体这一区域的CEPH YACs，但没有成功。然而，我们使用来自D16S125-D16S84区间的15个不同探针筛选了一个排列的P1文库，并鉴定了56个独立的P1克隆。间隔时间内只有一个探针无法识别P1克隆。根据限制性内切酶分析确定了44个P1克隆的独特性，其中42个克隆来自16p13.3染色体，基于FISH到中期染色体。700-kb的区间可以由一个由12个P1克隆和1个cosmid克隆组成的序列就绪contig来定义。我们的研究支持使用多个文库来生成定位克隆所需的物理试剂，并鼓励使用基于大肠杆菌的大插入克隆系统来恢复yac缺陷染色体间隔的克隆。
As part of an effort to identify the gene responsible for the predominant form of polycystic kidney disease (PKD1), we used a gridded human P1 library for contig assembly. The interval of interest, a 700-kb segment on chromosome 16p13.3, can be physically delineated by the genetic markers D16S125 and D16S84 and chromosomally characterized as a GC-rich isochore enriched for CpG islands, genes, and Alu-like repeats. Our attempts to recover CEPH YACs that encode this region of chromosome 16 were unsuccessful. However, we screened an arrayed P1 library using 15 distinct probes from the D16S125-D16S84 interval and identified 56 independent P1 clones. Only one probe from the interval was unsuccessful in identifying a P1 clone. Forty-four P1 clones were determined to be unique based on restriction enzyme analysis, and 42 of these were found to originate from chromosome 16p13.3, based on FISH to metaphase chromosomes. The 700-kb interval could be defined by a single sequence-ready contig comprised of 12 P1 clones and 1 cosmid clone. Our studies support the use of multiple libraries to generate the requisite physical reagents for positional cloning and encourage the use of Escherichia coli-based large-insert cloning systems to recover clones from YAC-deficient chromosomal intervals.