Evaluation of heat shock protein 70 as a biomarker of environmental stress in Fucus serratus and Lemna minor
Evaluation of heat shock protein 70 as a biomarker of environmental stress in Fucus serratus and Lemna minor
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DOI：
10.1080/13547500410001732610
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发表时间：
2004-03-01
期刊：
影响因子：
2.6
通讯作者：
Williams, JHH
中科院分区：
文献类型：
--
作者：
Ireland, HE;Harding, SJ;Williams, JHH
已知热休克蛋白（Hsps）在响应短期应激时被诱导。本研究的目的是评估热休克蛋白70作为生物标志物的压力所产生的温度升高，渗透压，暴露于镉和氯化钠在海洋大型藻类和淡水植物物种。间接竞争酶联免疫吸附试验（IC-ELISA）的工作范围为0.025 - 10 μ g/ml（-1），使用单克隆抗体提出了对纯化的热休克蛋白70的菜豆（绿豆）。将墨角藻（齿纹藻）、角叉菜（Stackhouse或角叉菜）、石莼（海莴苣）和浮萍（普通浮萍）样品提取物强制降解长达24小时，然后在IC-ELISA中进行检测。Western blot证实了Hsp70的存在和单克隆抗体的交叉反应性。热休克反应，确认在每个物种使用2小时42 degreesC处理。热休克后，Hsp70浓度增加，在2小时达到峰值（F。serratus）或4h（L.小，之后浓度下降。渗透胁迫和镉胁迫也导致F. serratus和L.与未强制降解对照相比，轻微。在这两种情况下，渗透胁迫和金属胁迫，热休克蛋白70的生产增加到最大值，随后下降的应激水平的增加。结果表明，热休克蛋白70 IC-ELISA可以潜在地应用于检测这些水生物种的压力，虽然它可能是最有效的结合使用时，与其他测量提供一个应激特异性生物标志物的档案或指纹。
Heat shock proteins ( Hsps) are known to be induced in response to short-term stress. The present study aimed to evaluate the potential of Hsp70 as a biomarker of stress produced by increased temperature, osmotic pressure, and exposure to cadmium and sodium chloride in marine macroalgae and fresh water plant species. An indirect competitive enzyme-linked immunosorbent assay (IC-ELISA) was developed with a working range of 0.025 - 10 mug ml(-1) using a monoclonal antibody raised against purified Hsp70 of Phaseolus aureus (mung bean). Fucus serratus ( toothed wrack), Chondrus crispus (Stackhouse or Carrageen moss), Ulva lactuca ( sea lettuce) and Lemna minor ( common duckweed) sample extracts were stressed for up to 24 h and then tested in the IC-ELISA. The presence of Hsp70 and cross-reactivity of the monoclonal antibody was confirmed by Western blot. The heat shock response was confirmed in each species using a 2-h 42degreesC treatment. Following heat shock, Hsp70 concentrations increased to a peak at 2 h ( F. serratus) or 4 h ( L. minor), after which concentrations decreased. Osmotic and cadmium stresses also resulted in elevated Hsp70 concentrations in samples of F. serratus and L. minor when compared with unstressed controls. In both, osmotic and metal stress, the production of Hsp70 increased to a maximum and subsequently decreased as the stressor levels increased. Results suggest that Hsp70 IC-ELISA could potentially be applied to the detection of stress in these aquatic species, although it would probably be most effective when used in conjunction with other measurements to provide a stressor-specific biomarker profile or fingerprint.