A lectin microarray approach for the rapid analysis of bacterial glycans
A lectin microarray approach for the rapid analysis of bacterial glycans
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DOI：
10.1038/nprot.2006.76
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发表时间：
2006-01-01
期刊：
影响因子：
14.8
通讯作者：
Mahal, Lara K.
中科院分区：
文献类型：
--
作者：
Hsu, Ku-Lung;Mahal, Lara K.
微生物细胞表面碳水化合物的快速评估对于了解细菌利用聚糖建立致病或共生关系的机制至关重要。微生物聚糖分析由于可能的碳水化合物结构的巨大多样性及其动态性质而变得复杂。细菌可以通过以相位可变的方式打开和关闭相关基因来快速改变其聚糖外壳。目前，缺乏适当的工具来研究动态碳水化合物的变化。在这里，我们提出了一个凝集素微阵列协议的细胞表面微生物糖的高通量评价。荧光细菌与这些阵列的结合模式提供了一种简单的方法，可以根据细菌表面的碳水化合物对其进行指纹识别。此外，该方法提供了一个快速，平行的评价聚糖从多个细菌样品，允许动态变化的碳水化合物结构进行研究。整个过程需要近似12小时，但微阵列的打印可以提前进行。
Rapid evaluation of microbial cell-surface carbohydrates is essential to understanding the mechanisms by which bacteria use glycans to establish pathogenic or symbiotic relationships. Microbial glycan analysis is complicated both by the vast diversity of possible carbohydrate structures and by their dynamic nature. Bacteria can rapidly alter their glycan coats by switching the genes that are involved on and off in a phase-variable manner. Currently, there is a lack of appropriate tools for studying dynamic carbohydrate alterations. Here, we present a lectin microarray protocol for the high-throughput evaluation of cell-surface microbial sugars. The binding patterns of fluorescent bacteria to these arrays provide a simple means to fingerprint bacteria based on their surface carbohydrates. In addition, this method provides a rapid, parallel evaluation of glycans from multiple bacterial samples, allowing dynamic changes in carbohydrate structures to be studied. The entire procedure takes similar to 12 h but the printing of the microarray can be performed in advance.