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Optimization of electrotransformation (ETF) conditions in lactic acid bacteria (LAB).
Journal of Microbiological Methods ( IF 2.2 ) Pub Date : 2020-05-14 , DOI: 10.1016/j.mimet.2020.105944
Chao Wang 1 , Yanhua Cui 1 , Xiaojun Qu 2
Affiliation  

Lactic acid bacteria (LAB) play important roles in the food industry, animal husbandry and medicine which are closely related to human life. Modern gene engineering technology is the major means to reveal gene functions, study metabolic pathways and metabolomes of strains and improve the properties of strains. However, up to now, the molecular technologies that can be applied to LAB are still very scarce. One of the main reasons for this phenomenon is the low efficiency of transformation, furthermore, the transformation protocols developed are strain specific. At present, the most common method in the transformation of LAB is electrotransformation (ETF), which has relatively high transformation efficiency, convenient operation and good reproducibility. In the process of ETF, many factors may be involved in the regulation of the ETF efficiency of the strains, including the characteristics of the strains, the properties of exogenous plasmids, the parameters of electric pulse, the application of cell wall weakening agents, the composition of washing and electroporation buffers, the resuspending culture media and heat treatment. In recent years, other methods have also been utilized to boost the transformation efficiency of strains, such as combined chemical-physical methods and bacterial conjugation. Various experimental parameters and alternative technologies to ETF which may impact the transformation efficiency are summarized in this paper. This review describes some meaningful factors for the development of transformation systems, and provides direction and foundations for the optimization and establishment of novel transformation strategies of LAB.

中文翻译:

优化乳酸菌(LAB)中的电转化(ETF)条件。

乳酸菌(LAB)在与人类生活密切相关的食品工业,畜牧业和医学中发挥着重要作用。现代基因工程技术是揭示基因功能,研究菌株的代谢途径和代谢组以及改善菌株特性的主要手段。但是,到目前为止,可用于LAB的分子技术仍然非常匮乏。造成这种现象的主要原因之一是转化效率低,此外,开发的转化方案是针对菌株的。目前,LAB转化最常用的方法是电转化(ETF),其转化效率较高,操作方便,重现性好。在ETF的过程中,菌株ETF效率的调节可能涉及许多因素,包括菌株的特性,外源质粒的特性,电脉冲的参数,细胞壁弱化剂的应用,洗涤和电穿孔缓冲液的组成,重新悬浮培养基和热处理。近年来,还采用了其他方法来提高菌株的转化效率,例如化学-物理方法和细菌结合方法。本文总结了可能影响转化效率的各种实验参数和ETF替代技术。这篇评论描述了开发转换系统的一些有意义的因素,
更新日期:2020-05-15
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