Colony hybridization to identify mammalian cells containing amplified, transfected, or expressed sequences.
Colony hybridization to identify mammalian cells containing amplified, transfected, or expressed sequences.
复制标题
集落杂交可鉴定含有扩增、转染或表达序列的哺乳动物细胞。
DOI：
10.1007/bf01534640
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发表时间：
1988
期刊：
影响因子：
--
通讯作者：
Rubin,LM
中科院分区：
文献类型：
--
作者：
Rossman,TG;Rubin,LM
变异细胞克隆的鉴定在阐明各种生化途径中发挥了重要作用。通常通过改变培养基或生长条件来选择这样的克隆，以使所需变体具有选择性优势。克隆DNA探针的可用性使得即使在没有选择的情况下也可以鉴定含有所需序列的细菌菌落或病毒噬斑。我们在这里描述了一种新的方法，用于识别和分离含有探针存在的任何序列的哺乳动物细胞。首先将哺乳动物细胞克隆复制到尼龙布上。原始克隆和复制品生长几天以扩大克隆。将原始培养皿上的所有克隆原位转移至滤器并进行杂交处理。通过改变处理，可以鉴定克隆中的DNA或RNA序列。当通过杂交定位感兴趣的克隆时，可以从尼龙复制品中分离细胞。我们期望这种方法在分离含有扩增、转染或突变基因的克隆以及那些表现出基因表达改变的克隆中具有价值。
The identification of variant cell clones has played an important role in the elucidation of various biochemical pathways. Such clones are typically selected by altering the medium or the growth conditions to give a selective advantage to the desired variant. The availability of cloned DNA probes has made it possible to identify bacterial colonies or viral plaques containing desired sequences, even in the absence of selection. We describe here a new method for the identification and isolation of mammalian cells containing any sequence for which a probe exists. Mammalian cell clones are first replicated onto nylon cloth. The original clones and the replicas are grown for a few days to enlarge the clones. All the clones on the original dish are transferred in situ to a filter and processed for hybridization. By altering the processing, either DNA or RNA sequences in the clones can be identified. When a clone of interest is located by hybridization, the cells can be isolated from the nylon replicas. We expect this method to be of value in the isolation of clones containing amplified, transfected, or mutated genes, as well as those exhibiting altered gene expression.