The first part of this project is inspired on a theoretical work that we have previously published about the physical constraints in the condensation of chromatin in metaphase chromosomes. The general objective of this part is the study of the higher order structures contained in the condensed forms of chromatin. We will develop two complementary approaches: a) study of the structures formed by the association of the 30-40 nm fibers; b) study of the higher order structures inside the metaphase chromosomes. We are planning to investigate the ionic conditions, compatible with the cellular values known at present, that are necessary to produce fiber association. We will prepare metaphase and prometaphase chromosomes from HeLa cells, and from species (chicken and \i Drosophila)\i0 having small chromosomes in their karyotype. To study the chromatin structures within chromosomes, we are planning to use ionic conditions, and treatments with denaturant agents, capable of producing a partial denaturation of the chromatids. In these studies we will use transmission and scanning electron microscopy, and atomic force microscopy. In the second part of the project we will carry out a detailed study of the peroxyoxalate chemiluminescent reaction, with the purpose to improve the sensitivity previously obtained in our laboratory in the detection of proteins and DNA on membranes. This is an important objective because this reaction shows a very high
|Effective start/end date||1/12/02 → 1/12/05|
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