Author/Editor		Vilfan, Tanja
Title		Uravnavanje s stresom inducibilne epimeraze glive Rhizopus nigricans
Type		monografija
Place		Ljubljana
Publisher		Medicinska fakulteta
Publication year		2001
Volume		str. 80
Language		slo
Abstract		In the filamentous fungus Rhizopus nigricans (R.nigricans) from phylum Zygomycota addition of different steroid hormones to the growth medium induces hydroxylating system, which presumably transforms steroid hormones into more soluble and therefore less toxic products. Besides inducing hydroxylation system of R. nigricans some steroid hormones provoke stress response during which the epimerase mRNA and also Hsp mRNA s are upregulated. Elevated levels of the epimerase mRNA were detected by Northern blot analyses after treatment of mycelia with progesterone, testosterone in deoxycorticosterone and also after exposure to other stresses like heat shock, osmotic and oxidative stress, starvation and exposure to some heavy metals. Since the stress response was so far studied only at the level of mRNA synthesis we tried to establish whether elevated levels of the epimerase mRNA are followed by a raise in protein synthesis. To test that we prepared mouse polyclonal antibodies against the recombinant polypeptide with a partial amino acid sequence of the epimerase. The recombinant polypeptide that we used as an antigen was prepared as a fusion protein with glutathione S-transferase. Mouse polyclonal antibodies against the recombinant polypeptide with a partial amino acid sequence of the epimerase were used for Western blot analyses. We were unable to demonstrate the connection between induced mRNA synthesis in mycelium after the exposure to osmotic stress, heat shock or steroid hormones, but elevated levels of epimerase were detected in protein samples from mycelium exposed to the induction buffer compared to the lower levels of the protein in protein samples from mycelium exposed to the induction buffer with glucose. (Abstract truncated at 2000 characters).
Descriptors		RHIZOPUS EPIMERASES RNA, MESSENGER PROMOTER REGIONS (GENETICS) ANTIBODIES PLASMIDS ESCHERICHIA COLI TRANSFORMATION, GENETIC BASE SEQUENCE DNA FRAGMENTATION RECOMBINANT PROTEINS