Yu J, Qi J, Kao F T
Eleanor Roosevelt Institute for Cancer Research, Denver, Colorado 80206.
Somat Cell Mol Genet. 1994 Mar;20(2):133-6. doi: 10.1007/BF02290682.
We report the construction and characterization of a region-specific microdissection library for human chromosome 2p11-p13. This library (designated 2P4 library) is large, comprising 600,000 recombinant microclones. Thirty to 40% of the clones contain unique sequences. The insert sizes range from 100 to 800 bp, with a mean of 380 bp. A subset of the microclones was selected, based on their weak or no hybridization to total human DNA, for further analysis. Of 50 single-copy microclones analyzed, 35 clones (70%) were derived from human and are chromosome 2-specific. The insert sizes and the hybridizing genomic HindIII fragments of these clones were also determined. The 2P4 microdissection library and the single-copy microclones from the library are useful in preparing STS (sequence-tagged site) to isolate corresponding YAC (yeast artificial chromosome) or other clones with large inserts and for isolating region-specific cDNA clones as candidate genes for cloning disease-related genes assigned to this region.
我们报告了人类染色体2p11 - p13区域特异性显微切割文库的构建及特性分析。该文库（命名为2P4文库）规模较大，包含600,000个重组微克隆。30%至40%的克隆含有独特序列。插入片段大小范围为100至800 bp，平均为380 bp。基于微克隆与总人类DNA的弱杂交或无杂交情况，选择了一部分微克隆进行进一步分析。在分析的50个单拷贝微克隆中，35个克隆（70%）来源于人类且是2号染色体特异性的。还确定了这些克隆的插入片段大小及杂交的基因组HindIII片段。2P4显微切割文库以及来自该文库的单拷贝微克隆可用于制备序列标签位点（STS）以分离相应的酵母人工染色体（YAC）或其他具有大插入片段的克隆，也可用于分离区域特异性cDNA克隆作为克隆定位于该区域的疾病相关基因的候选基因。
