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Flourescence-based isolation of differentially induced genes

机译:基于荧光的差异诱导基因分离

摘要

Regulatory elements (e.g. promoters) activated by a stimulus are isolated by a FACS-based method. Preferably, a library of random fragments representative of a target (e.g. bacterial) genome are cloned in front of a promoterless gfp (green fluorescent protein) sequence in a plasmid, and inserted into target cells. The resulting target cell mixture is sorted according to GFP levels in the presence and the absence of the stimulus. Suitable stimuli include compounds of interest (e.g. drugs), environmental factors (e.g. extracellular acidity), and complex stimuli such as in vivo environments of hosts infected by the target cells. The method allows identifying pathogen genes which are selectively expressed during infection.
机译:通过基于FACS的方法分离被刺激激活的调节元件(例如启动子)。优选地,将代表靶标(例如细菌)基因组的随机片段文库克隆到质粒中无启动子的gfp(绿色荧光蛋白)序列的前面,并插入靶标细胞中。在存在和不存在刺激的情况下,根据GFP水平对得到的靶细胞混合物进行分类。合适的刺激物包括目标化合物(例如药物),环境因素(例如细胞外酸度)和复杂的刺激物,例如被靶细胞感染的宿主的体内环境。该方法允许鉴定在感染期间选择性表达的病原体基因。

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