通过对cDNA文库进行差异显示筛选来分离发育调控基因。
Isolation of developmentally regulated genes by differential display screening of cDNA libraries.
作者信息
Gupta R, Thomas P, Beddington R S, Rigby P W
机构信息
Division of Eukaryotic Molecular Genetics and Division of Mammalian Development,MRC National Institute for Medical Research, The Ridgeway, Mill Hill, London NW7 1AA, UK.
出版信息
Nucleic Acids Res. 1998 Oct 1;26(19):4538-9. doi: 10.1093/nar/26.19.4538.
mRNA differential display RT-PCR has been extensively used for the isolation of genes differentially expressed between RNA populations. We have assessed its utility for the identification of developmentally regulated genes in plasmid cDNA libraries derived from individual tissues dissected from early mouse embryos. Using plasmid Southern blot hybridisation as a secondary screen, we are able to identify such genes and show by whole-mount in situ hybridisation that their expression pattern is that expected from the differential display profile.
mRNA差异显示逆转录聚合酶链反应已被广泛用于分离RNA群体之间差异表达的基因。我们评估了它在从早期小鼠胚胎解剖的单个组织衍生的质粒cDNA文库中鉴定发育调控基因的效用。使用质粒Southern印迹杂交作为二级筛选，我们能够鉴定出此类基因，并通过整体原位杂交表明它们的表达模式与差异显示图谱预期的一致。
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