Immunological Detection of UV Induced Cyclobutane Pyrimidine Dimers and (6-4) Photoproducts in DNA from Reference Bacteria and Natural Aquatic Populations
Kraft, S. / Obst, U. / Schwartz, T. (2011)
Journal of Microbiological Methods 84 (2011), 3, 435-441
- Date: 2011
Kraft, S. / Obst, U. / Schwartz, T. (2011): „Immunological Detection of UV Induced Cyclobutane Pyrimidine Dimers and (6-4) Photoproducts in DNA from Reference Bacteria and Natural Aquatic Populations“. In: Journal of Microbiological Methods 84 (2011), 3, 435-441
UV light-caused DNA damage is a widespread field of study. As UV light has the biological effect of inactivating or killing bacteria, it is used for water disinfection. Due to this application, it is important to study the DNA damage efficiencies and regeneration capacities in bacteria after UV irradiation.
Two monoclonal antibodies, anti-CPD and anti-(6–4) PP, were applied for an immunoassay of UV-induced DNA modifications. Cyclobutane pyrimidine dimer (CPD) and 6–4 photoproduct (6–4 PP) were detected in the reference bacteria Pseudomonas aeruginosa and Enterococcus faecium, and in natural water communities.
The antibody-mediated detection signals increased with the UV doses from 100–400 J/m2. Here, the CPD-specific signals were stronger than the (6–4) PP-specific signals. These immunological results were in accordance with parallel cultivation experiments. All UV-irradiated bacteria showed a reduction of their growth rate depending on UV application by several orders of magnitudes.
The immunoassay was also applied to three types of natural aquatic habitats with different cell densities. Besides artificial UV irradiation, it was possible to visualize natural sunlight effects on these natural bacterial communities. Light-dependent and dark repair processes were distinguished using the established immunological assays. The antibody-mediated analyses presented are fast methods to detect UV-induced DNA lesions and repair capacities in selected bacterial species as well as in entire natural mixed populations.