Recently, we have published the results of our research on the structure of small chromatin fragments of chicken erythrocyte (Bartolomé et al., J. Cell. Sci., 107 (1994) 2983-2992 y J. Biol. Chem., 270 (1995) 22514-22521). In this research we have used nondenaturing gel electrophoresis and electron microscopy techniques, which have allowed us to study the folding and internal structure of these small chromatin fragments. In this project we are planning to apply the electrophoretic and electron microscopy procedures used for chicken erythrocytes, to study of the estructure of small chromatin fragments corresponding to tissues with different levels of genetic activity (rat liver, calf thymus, sea urchin sperm). We will study yeast in order to examine whether a system without histone H1 can produce folded chromatin. Furthermore, in order to increase our knowledge about the internal structure of chromatin fibers, we will try ro find conditions for the progressive denaturation of the compact chromatin fragments. On the other hand, in our laboratory we have developed fluorescent methods for the detection of proteins in electrophoretic gels [Dabán et al., \i Anal. Biochem.\i0 \i 199\i0 (1991) 169-174; Bermúdez et al., \i BioTechniques\i0 , \i 16\i0 (1994) 621-624]. These contributions, and all our previous experience in the field of the fluorescent probes, have stimulated us to investigate the applications in Molecular Biology of chemiluminescent reagents (oxalic acid esters) that are able to excite different fluorophores. In the secon part of the project we are planning to develop methods for the use the chemiluminescent reagents TCPO and DNPO for the detection of proteins (in gels and Western blots) and DNA (in current and sequencing gels, and in Southern blots).
|Effective start/end date||1/11/96 → 1/11/99|
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