Plant-based FRET biosensor discriminates environmental zinc levels

被引:9
|
作者
Adams, Joshua P. [1 ]
Adeli, Ardeshir [2 ]
Hsu, Chuan-Yu [3 ]
Harkess, Richard L. [4 ]
Page, Grier P. [5 ]
dePamphilis, Claude W. [6 ]
Schultz, Emily B. [3 ]
Yuceer, Cetin [3 ]
机构
[1] Univ Arkansas Monticello, Sch Forest Resources, Monticello, AR USA
[2] ARS, USDA, Mississippi State, MS USA
[3] Mississippi State Univ, Dept Forestry, Mississippi State, MS 39762 USA
[4] Mississippi State Univ, Dept Plant & Soil Sci, Mississippi State, MS 39762 USA
[5] RTI Int, Atlanta, GA USA
[6] Penn State Univ, Dept Biol, University Pk, PA 16802 USA
关键词
zinc interaction; environmental monitoring; ZIP transporter; ECFP; DsRED; ENERGY-TRANSFER BRET; RED FLUORESCENT PROTEIN; REAL-TIME DETECTION; ZIP FAMILY; EXPRESSION; GLUCOSE; POPLAR; GENES; TRANSPORT; CADMIUM;
D O I
10.1111/j.1467-7652.2011.00656.x
中图分类号
Q81 [生物工程学(生物技术)]; Q93 [微生物学];
学科分类号
071005 ; 0836 ; 090102 ; 100705 ;
摘要
Heavy metal accumulation in the environment poses great risks to flora and fauna. However, monitoring sites prone to accumulation poses scale and economic challenges. In this study, we present and test a method for monitoring these sites using fluorescent resonance energy transfer (FRET) change in response to zinc (Zn) accumulation in plants as a proxy for environmental health. We modified a plant Zn transport protein by adding flanking fluorescent proteins (FPs) and deploying the construct into two different species. In Arabidopsis thaliana, FRET was monitored by a confocal microscope and had a 1.4-fold increase in intensity as the metal concentration increased. This led to a 16.7% overall error-rate when discriminating between a control (1 mu m Zn) and high (10 mm Zn) treatment after 96 h. The second host plant (Populus tremula x Populu salba) also had greater FRET values (1.3-fold increase) when exposed to the higher concentration of Zn, while overall error-rates were greater at 22.4%. These results indicate that as plants accumulate Zn, protein conformational changes occur in response to Zn causing differing interaction between FPs. This results in greater FRET values when exposed to greater amounts of Zn and monitored with appropriate light sources and filters. We also demonstrate how this construct can be moved into different host plants effectively including one tree species. This chimeric protein potentially offers a method for monitoring large areas of land for Zn accumulation, is transferable among species, and could be modified to monitor other specific heavy metals that pose environmental risks.
引用
收藏
页码:207 / 216
页数:10
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