A molecular strategy designed for the rapid screening of gene traps based on sequence identity and gene expression pattern in adult mice.
A molecular strategy designed for the rapid screening of gene traps based on sequence identity and gene expression pattern in adult mice.
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一种分子策略，旨在根据成年小鼠的序列同一性和基因表达模式快速筛选基因陷阱。
DOI：
10.1023/a:1018465402294
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发表时间：
1997
影响因子：
3
通讯作者：
Mark,WH
中科院分区：
文献类型：
--
作者：
Holzschu,D;Lapierre,L;Neubaum,D;Mark,WH
我们设计了一种基于 DNA 序列信息和体外基因表达分析快速筛选小鼠胚胎干 (ES) 细胞中基因陷阱的策略。在初步鉴定表达 β-半乳糖苷酶的 ES 细胞克隆后，立即对标记的 RNA 转录物进行克隆和测序以确定其身份。发现的新基因序列被用来探测 Northern blots，以检查其同源基因的表达模式。我们对 30 个 cDNA 克隆的初步表征表明，超过一半的标记序列是新的小鼠基因，其中 40% 在成年小鼠组织中显示出受限的表达模式。这种基因陷阱的分子表征快速、可靠，非常适合哺乳动物发育基因的大规模筛选。此外，由于基因陷阱插入经常破坏标记的宿主基因，因此 ES 细胞可用于生产转基因动物，用于基因功能的遗传分析
We have devised a strategy to rapidly screen gene traps in mouse embryonic stem (ES) cells based on DNA sequence information and an in vitro analysis of gene expression. After the initial identification of ES cell clones expressing β-galactosidase, tagged RNA transcripts were immediately cloned and sequenced in order to determine their identities. Novel gene sequences found were used to probe northern blots to examine the expression patterns of their cognate genes. Our initial characterization of 30 cDNA clones indicated that more than half of the tagged sequences were novel mouse genes and of these 40% showed a restricted pattern of expression in adult mouse tissues. This molecular characterization of gene traps is quick, reliable and well suited for the large-scale screening of mammalian developmental genes. Furthermore, since gene trap insertion frequently disrupts the tagged host gene, the ES cells can be used to produce transgenic animals for a genetic analysis of gene function