This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. The investigators have developed new techniques that permit the retention of protein antigens in biological materials throughout their preparation for TEM. This procedure avoids oxidation and extraction of proteins which routinely occur with conventional embedding protocols. The workers wish to test the methodology on polytene chromosomes of Drosophila spp., where proteins of general interest are to be found. These reflect the highly conserved functions of chromatin packing in the centromere and telomeres, as well as in portions of the genome that are transcriptionally inactivated through development. The chromatin-attached proteins HP1 and Pc will be localized by the principal investigator's technology. Electron tomography and reconstruction of the 3-dimensional image datasets are to be done at NCMIR.
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