| 摘要 |
Quantitatively evaluating the global and specific capacities for DNA repair in a biological medium, is new. Process for quantitative evaluation of the global and specific capacities for DNA repair in a biological medium comprises: (a) preparing a series of plasmids (P), each containing distinct DNA lesions, by independently treating them with a physical and/or chemical agent and recovery of the supercoiled fraction; (b) characterizing the lesions in each P; (c) depositing P, and at least one control supercoiled plasmid without lesions, on a single solid support, according to a pre-established scheme, to produce a functional support divided into x distinct zones (x = number of media to be tested simultaneously) where each zone contains the set of P; (d) incubating the support with different repair solutions, each containing the enzyme activities required for repair, ATP (adenosine triphosphate) and a system for regenerating it, a labeled nucleotide triphosphate and other components required for enzymatic activity, at 30 degrees Centigrade for 1-5, preferably 3, hours, with each repair solution being applied to a different zone A1-Ax; (e) washing the support; (f) (in)direct measurement of the signal produced by the label, incorporated into DNA during the repair reaction, in each zone; (g) recording and quantifying the signal from each plasmid in each zone; and (h) determining the ratio of signal from each plasmid relative to that from the control. |
