A Convenient Fluorescence-Based Assay for the Detection of Sucrose Transport and the Introduction of a Sucrose Transporter from Potato into Clostridium Strains
A Convenient Fluorescence-Based Assay for the Detection of Sucrose Transport and the Introduction of a Sucrose Transporter from Potato into Clostridium Strains
复制标题
一种方便的基于荧光的蔗糖转运检测方法以及将马铃薯蔗糖转运蛋白引入梭菌菌株中
DOI：
10.3390/molecules24193495
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发表时间：
2019-10-01
期刊：
影响因子：
4.6
通讯作者：
Du, Liqin
中科院分区：
文献类型：
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作者：
Zhang, Zhikai;Lin, Lihua;Du, Liqin
对于梭菌菌株，需要一种便捷有效的蔗糖转运检测方法。传统方法，比如碳 - 14 - 蔗糖同位素标记法，使用放射性物质，对很多实验室来说并不方便。在此，将一种来自马铃薯的蔗糖转运蛋白引入梭菌，并利用基于七叶苷的荧光检测法来分析梭菌菌株中的蔗糖转运情况。这表明异源表达的马铃薯蔗糖转运蛋白在梭菌中具有功能。高水平蔗糖转运的重组工程将有助于梭菌菌株中的蔗糖发酵。本文所述的检测方法为研究蔗糖转运蛋白在梭菌中异源表达后的功能提供了一个重要的技术平台。
A convenient and effective sucrose transport assay for Clostridium strains is needed. Traditional methods, such as 14C-sucrose isotope labelling, use radioactive materials and are not convenient for many laboratories. Here, a sucrose transporter from potato was introduced into Clostridium, and a fluorescence assay based on esculin was used for the analysis of sucrose transport in Clostridium strains. This showed that the heterologously expressed potato sucrose transporter is functional in Clostridium. Recombinant engineering of high-level sucrose transport would aid sucrose fermentation in Clostridium strains. The assay described herein provides an important technological platform for studying sucrose transporter function following heterologous expression in Clostridium.