Ripp S, Jegier P, Birmele M, Johnson C M, Daumer K A, Garland J L, Sayler G S
Center for Environmental Biotechnology, University of Tennessee, Knoxville, TN 37996-1605, USA.
J Appl Microbiol. 2006 Mar;100(3):488-99. doi: 10.1111/j.1365-2672.2005.02828.x.
To incorporate into the lambda phage genome, a luxI-based acyl-homoserine lactone (AHL) synthase genetic construct and exploit the autoamplified power of quorum sensing to translate a phage infection event into a chemical signature detectable by a lux-based bioluminescent bioreporter, with focus towards facile detection of microbial pathogens.
The luxI gene from Vibrio fischeri was inserted into the lambda phage genome to construct a model phage-based biosensor system for the general detection of Escherichia coli. The AHL signalling molecules synthesized upon phage infection are detected by an AHL-specific bioluminescent bioreporter based on the luxCDABE gene cassette of V. fischeri. The assay generates target-specific visible light signals with no requisite addition of extraneous substrate. This binary reporter system was able to autonomously respond to lambda phage infection events at target E. coli concentrations ranging from 1 x 10(8) to 1 CFU ml(-1) within 1.5-10.3 h, respectively, in pure culture. When assayed against artificially contaminated lettuce leaf washings, detection within an E. coli inoculum range from 1 x 10(8) to 130 CFU ml(-1) was achieved within 2.6-22.4 h, respectively.
The initial feasibility of binary phage-based reporter assays indicates that quorum sensing can be used to translate a phage infection event into an autoamplified chemical signature.
With further modification, binary phage-based reporter assays may be capable of rapidly and cost effectively detecting pathogenic agents at very low population densities.
将基于luxI的酰基高丝氨酸内酯（AHL）合酶基因构建体整合到λ噬菌体基因组中，并利用群体感应的自动放大能力，将噬菌体感染事件转化为基于lux的生物发光生物报告器可检测的化学信号，重点是便于检测微生物病原体。
将费氏弧菌的luxI基因插入λ噬菌体基因组，构建用于大肠杆菌通用检测的基于噬菌体的模型生物传感器系统。基于费氏弧菌的luxCDABE基因盒的AHL特异性生物发光生物报告器可检测噬菌体感染时合成的AHL信号分子。该检测无需添加额外底物即可产生目标特异性可见光信号。在纯培养中，该二元报告系统能够在1.5 - 10.3小时内，分别对浓度范围为1×10⁸至1 CFU ml⁻¹的目标大肠杆菌中的λ噬菌体感染事件自主做出反应。当针对人工污染的生菜叶洗涤液进行检测时，分别在2.6 - 22.4小时内实现了对浓度范围为1×10⁸至130 CFU ml⁻¹的大肠杆菌接种物的检测。
基于噬菌体的二元报告检测的初步可行性表明，群体感应可用于将噬菌体感染事件转化为自动放大的化学信号。
经过进一步改进，基于噬菌体的二元报告检测可能能够在极低的种群密度下快速且经济高效地检测病原体。
