Cross-hybridization of closely related Genes on high-density macroarrays
Cross-hybridization of closely related Genes on high-density macroarrays
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DOI：
10.2144/02323pf01
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发表时间：
2002-03-01
期刊：
影响因子：
2.7
通讯作者：
LaBrie, ST
中科院分区：
文献类型：
--
作者：
Miller, NA;Gong, Q;LaBrie, ST
DNA宏阵列用于分子生物学研究的许多领域，其应用范围从基因发现到基因表达谱分析。随着越来越多的含有功能或途径相关基因的专用宏阵列变得可用，需要解决的问题是可以实现的高度相似基因之间的杂交信号特异性水平。我们检测了LifeGrid（TM）宏阵列区分密切相关基因间杂交信号的能力，确定了序列同一性在52%到94%之间的基因间交叉杂交水平。来自五个蛋白质家族的基因片段排列在尼龙过滤器上。随后将滤膜与P-33标记的探针杂交，所述探针是从含有每个蛋白质家族的代表的合成mRNA转录物的池制备的。我们发现，含有具有94%序列同一性的片段显示相对较少的交叉杂交。我们的结论是，这个宏阵列系统是非常具体的，从密切相关的基因的杂交信号可以可靠地测量。
DNA macroarrays are used in many areas of molecular biology research for applications ranging from gene discovery to gene expression profiling. As an increasing number of specialized macroarrays containing genes related by function or pathway are becoming available, a question that needs to be addressed is the level of hybridization signal specificity between highly similar genes that can be achieved. We have examined the ability of our LifeGrid(TM) macroarrays to distinguish hybridization signals between closely related genes.We determined the level of cross-hybridization among genes ranging front 52% to 94% sequence identity. Fragments of genes from five protein families were arrayed onto nylon filters. The filters were subsequently hybridized with a P-33-labeled probe prepared from a pool of synthetic mRNA transcripts containing a representative of each protein family, We found that fragments containing sequences with tip to 94% sequence identity displayed relatively little cross-hybridization. We conclude that this macroarray system is very specific and that hybridization signals from closely related genes can be reliably measured.