Avtor/Urednik     Černila, Boštjan
Naslov     Geni stresnih proteinov iz družine hsp7- v glivi Rhizopus nigricans in dejavniki, ki vplivajo na njihovo izražanje
Tip     monografija
Kraj izdaje     Ljubljana
Založnik     Medicinska fakulteta
Leto izdaje     2001
Obseg     str. 111
Jezik     slo
Abstrakt     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 (Černila 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 as a 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 mRNA level at different stress conditions, since expression of this subgroup of hsp70s is most commonly up-regulated with different stressors. We found that all three cytosolic hsp70s from R. nigricans had similar patterns of expression. (Abstract truncated at 2000 characters).
Deskriptorji     RHIZOPUS
HEAT-SHOCK PROTEINS 70
GENE EXPRESSION
DNA PROBES
SEQUENCE ANALYSIS, DNA
BASE SEQUENCE
PROMOTER REGIONS (GENETICS)
ESCHERICHIA COLI
DNA, RECOMBINANT