Expression in Escherichia coli, purification, refolding and antifungal activity of an osmotin from Solanum nigrum

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Expression in , purification, refolding and antifungal activity of an osmotin from -1

Ed at 37°C for 3 h and induced by 0.4 mM IPTG (when indicated). . Molecular mass marker (LMW, Amersham Pharmacia Biotech); . Total protein fraction from non-induced culture; . Total protein fraction from IPTG-induced culture; . Soluble protein fraction from IPTG-induced culture; . Insoluble protein fraction from IPTG-induced culture. . Western blot analysis of expressed His-tagged mature SnOLP probed with His-monoclonal antibody. . Soluble protein fraction from IPTG-induced culture; . Insoluble protein fraction from IPTG-induced culture. . SDS-PAGE analysis of insoluble His-tagged mature SnOLP which was urea solubilized and subsequently purified by immobilized-metal (Ni) affinity chromatography (IMAC). and . Eluates from IMAC. In . and . the gels were stained with Coomassie brilliant blue. His-tagged mature SnOLP protein is indicated by arrow heads.<p><b>Copyright information:</b></p><p>Taken from "Expression in , purification, refolding and antifungal activity of an osmotin from "</p><p>http://www.microbialcellfactories.com/content/7/1/7</p><p>Microbial Cell Factories 2008;7():7-7.</p><p>Published online 11 Mar 2008</p><p>PMCID:PMC2362109.</p><p></p

Expression in , purification, refolding and antifungal activity of an osmotin from -2

proteins were placed on plates 3 days after inoculation with fungal mycelia (plugs). Petri dishes were incubated at 29°C during the entire bioassay. Inhibitory effects of purified and renatured His-tagged mature SnOLP upon the fungi are observed as areas lacking mycelial growth. . var. ; 1 day after adding protein (a.a.p.); . , 5 days a.a.p. . Corresponds to the doses of 1 μg, 2 μg and 3 μg of purified and renatured His-tagged mature SnOLP, or to the concentrations of 0,1 μg/μL, 0,2 μg/μL and 0,3 μg/μL, respectively; . Corresponds to the dose of 10 μg of Bovine Serum Albumin (BSA), or to the concentration of 1 μg/μL; . Corresponds to the dose of 2000 U Nistatin, or to the concentration of 200 U/μL. . Average area (mm) of mycelial growth inhibition caused by renatured His-tagged mature SnOLP, as measured (software UTHSCSA Image Tool, Version 3.00 [57]) in three replicates, similar to the bioassay Petri dishes shown in A and B, for each fungus/oomycete and for each dose/concentration of SnOLP separately. Standard deviation bars are shown for each average column. The averages were statistically compared by using ANOVA and Tukey Test at the probability level of 1% (software Genes [58]). Different letters above the average columns (i.e. a, b and c), indicate that the average values were considered to be statistically different among each other, whereas statistically identical average values are indicated by the same letter. . var. ; 1 day after adding protein (a.a.p.); . , 5 days a.a.p. ; . , 12 days a.a.p.; . var. , 5 days a.a.p.; . f. sp. , 5 days a.a.p.<p><b>Copyright information:</b></p><p>Taken from "Expression in , purification, refolding and antifungal activity of an osmotin from "</p><p>http://www.microbialcellfactories.com/content/7/1/7</p><p>Microbial Cell Factories 2008;7():7-7.</p><p>Published online 11 Mar 2008</p><p>PMCID:PMC2362109.</p><p></p

Expression in , purification, refolding and antifungal activity of an osmotin from -4

Ed at 37°C for 3 h and induced by 0.4 mM IPTG (when indicated). . Molecular mass marker (LMW, Amersham Pharmacia Biotech); . Total protein fraction from non-induced culture; . Total protein fraction from IPTG-induced culture; . Soluble protein fraction from IPTG-induced culture; . Insoluble protein fraction from IPTG-induced culture. . Western blot analysis of expressed His-tagged mature SnOLP probed with His-monoclonal antibody. . Soluble protein fraction from IPTG-induced culture; . Insoluble protein fraction from IPTG-induced culture. . SDS-PAGE analysis of insoluble His-tagged mature SnOLP which was urea solubilized and subsequently purified by immobilized-metal (Ni) affinity chromatography (IMAC). and . Eluates from IMAC. In . and . the gels were stained with Coomassie brilliant blue. His-tagged mature SnOLP protein is indicated by arrow heads.<p><b>Copyright information:</b></p><p>Taken from "Expression in , purification, refolding and antifungal activity of an osmotin from "</p><p>http://www.microbialcellfactories.com/content/7/1/7</p><p>Microbial Cell Factories 2008;7():7-7.</p><p>Published online 11 Mar 2008</p><p>PMCID:PMC2362109.</p><p></p

Expression in , purification, refolding and antifungal activity of an osmotin from -0

To generate fragments coding for mature SnOLP form are shown. The orientations of the primers and , including the respective restriction cloning sites, are indicated by arrows. The His-tag encoded by the pQE30 vector is indicated. , Signal Peptide; , Carboxy-Terminal propeptide; , Methionine; *, Stop codon.<p><b>Copyright information:</b></p><p>Taken from "Expression in , purification, refolding and antifungal activity of an osmotin from "</p><p>http://www.microbialcellfactories.com/content/7/1/7</p><p>Microbial Cell Factories 2008;7():7-7.</p><p>Published online 11 Mar 2008</p><p>PMCID:PMC2362109.</p><p></p