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Geni stresnih proteinov iz družine hsp70 v glivi Rhizopus nigricans in dejavniki, ki vplivajo na njihovo izražanje : doktorska disertacija = Genes encodings stress proteins from hsp70 family in fungus Rhizopus nigricans and factors that influence their expression : doctoral thesis
Černila, Boštjan
The hsp70 multigene family of stress proteins is an evolutionary conserved group of molecular chaperones with diverse cellular functions. The regulation of hsp70 gene expression is very complex and ...still not completely understood. Since, so, far, the only hsp70s described in fungal phylum Zygomycota were at cDNA level in Rhizopus nigricans (^ernila at al.1999), we decided to further investigate the genes for these proteins. We constructed the R. nigricans genomic library in a bacteriophage vector. Following the screening of this library with radiolabeled cDNA fragment, encoding a conserved part of hsp70 asa probe, five genomic clones (G8, G12, G16, G17 and 702B) were selected. Comparison of nucleotide sequences of isolated genomic clones with the sequences of previously described cDNAs encoding three cytosolic hsp70 (hsp70-1, hsp70-2 and hsp70-3) had revealed that genomic clones G17, 702B and G16 corresponded to hsp70-1, hsp70-2 and hsp70-3, respectively, while genomic clones G8 and G12 encoded some new proteins. By searching protein databases for homologous proteins, we had found that clone G8 contained partial coding region for a protein highly homologous to a hsp70 associated with ribosomes, while clone G12 encoded a part of a protein homologous to hsp70 from endoplasmic reticulum. Southern blot analysis of R. nigricans genomic DNA showed that we had isolated all members of hsp70 family except mitochondrial hsp70. Furthermore, we investigated the expression of cytosolic hsp70s on mRNAlevel at different stress conditions, since expression of this subgroup ofhsp70s is most commonly up-regulated with different stressors. We found thatall three cytosolic hsp70s from R. nigricans had similar patterns of expression. (Abstract truncated at 2000 characters).
Type of material - dissertation
Publication and manufacture - Ljubljana : [B. Černila], 2001
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