Utility of LOV-based Fluorescent Reporters To Monitor the Real-Time Secretion and Localisation of Bacterial Effector Proteins

利用基于 LOV 的荧光报告基因监测细菌效应蛋白的实时分泌和定位

• 批准号: BB/H023518/1
• 负责人: Andrew Roe
• 金额: $ 12.32万
• 依托单位: University of Glasgow
• 依托单位国家: 英国
• 项目类别: Research Grant
• 财政年份: 2011
• 资助国家: 英国
• 起止时间: 2011 至 无数据
• 项目状态: 已结题
• 来源: https://gtr.ukri.org/projects?ref=BB%2FH023518%2F1
• 关键词: Utility; LOV; based; Fluorescent; Reporters

The project aims to develop and test a novel fluorescent marker that we have been working on. This is called LOV, which stands for 'Light, Oxygen, Voltage'. We work on a form of LOV which emits a strong green fluorescence upon blue light excitation. We believe that LOV might act as an alternative to green fluorescent protein (GFP), used extensively in biology for many applications including the study of protein folding, protein transport, RNA dynamics and responses to infection. In this project we will test to see if LOV can be used to 'tag' proteins that are made by bacteria and then injected into host cells by a dedicated secretion system. The project is strengthened by using expertise from 3 different lead scientists: Dr Chrisitie is a biochemist who has led research on the application of LOV in novel areas of biology, Dr Roe is a microbiologist working on gene regulation and bacterial-host cell interactions and Dr Enninga, a named collaborator, is a specialist in imaging bacterial proteins.

该项目旨在开发和测试我们一直在研究的一种新型荧光标记物。这就是LOV，代表“光，氧，电压”。我们研究了一种在蓝光激发下发出强烈绿色荧光的LOV。我们相信LOV可能作为绿色荧光蛋白（GFP）的替代品，广泛用于生物学中的许多应用，包括蛋白质折叠，蛋白质转运，RNA动力学和感染反应的研究。在这个项目中，我们将测试LOV是否可以用来“标记”由细菌制造的蛋白质，然后通过专用的分泌系统注入宿主细胞。该项目通过使用3位不同的首席科学家的专业知识得到加强：Chrisitie博士是一位生物化学家，他领导了LOV在生物学新领域的应用研究，Roe博士是一位微生物学家，致力于基因调控和细菌-宿主细胞相互作用，Enninga博士是一位命名的合作者，是细菌蛋白质成像专家。

项目成果

Tracking elusive cargo: Illuminating spatio-temporal Type 3 effector protein dynamics using reporters.

• DOI: 10.1111/cmi.12797
• 发表时间: 2018-01
• 期刊: Cellular microbiology
• 影响因子: 3.4
• 作者: O'Boyle N;Connolly JPR;Roe AJ
• 通讯作者: Roe AJ

Visualizing the Translocation and Localization of Bacterial Type III Effector Proteins by Using a Genetically Encoded Reporter System.

• DOI: 10.1128/aem.03418-15
• 发表时间: 2016-05
• 期刊: Applied and environmental microbiology
• 影响因子: 4.4
• 作者: Gawthorne JA;Audry L;McQuitty C;Dean P;Christie JM;Enninga J;Roe AJ
• 通讯作者: Roe AJ

Expression and regulation of the Escherichia coli O157:H7 effector proteins NleH1 and NleH2.

• DOI: 10.1371/journal.pone.0033408
• 发表时间: 2012
• 期刊: PloS one
• 影响因子: 3.7
• 作者: Holmes A;Lindestam Arlehamn CS;Wang D;Mitchell TJ;Evans TJ;Roe AJ
• 通讯作者: Roe AJ

Lighting Up Clostridium Difficile: Reporting Gene Expression Using Fluorescent Lov Domains.

• DOI: 10.1038/srep23463
• 发表时间: 2016-03-21
• 期刊: Scientific reports
• 影响因子: 4.6
• 作者: Buckley AM;Jukes C;Candlish D;Irvine JJ;Spencer J;Fagan RP;Roe AJ;Christie JM;Fairweather NF;Douce GR
• 通讯作者: Douce GR