Journal: Applied and Environmental Microbiology

Article Title: Bioprospecting for Trichothecene 3- O -Acetyltransferases in the Fungal Genus Fusarium Yields Functional Enzymes with Different Abilities To Modify the Mycotoxin Deoxynivalenol ▿ †

doi: 10.1128/AEM.01738-10

Figure Lengend Snippet: Sequences of primers used to amplify all or portions of TRI101/TRI201 and FsTRI12 a

Article Snippet: The TRI101/TRI201 proteins were diluted to 0.2 mg/ml in a degassed buffer containing 100 mM potassium phosphate at pH 8, and spectra were recorded from 190 to 260 nm in a model 202SF circular dichroism spectrophotometer (Aviv Biomedical).

Techniques: Sequencing, Clone Assay, Isolation

Journal: Applied and Environmental Microbiology

Article Title: Bioprospecting for Trichothecene 3- O -Acetyltransferases in the Fungal Genus Fusarium Yields Functional Enzymes with Different Abilities To Modify the Mycotoxin Deoxynivalenol ▿ †

doi: 10.1128/AEM.01738-10

Figure Lengend Snippet: Phylogenetic tree based on a total of 688 bp of sequence internal to the 3′ end of TRI101/TRI201. Sequences included in the tree are from a total of 54 strains of Fusarium: 33 strains from the Leslie collection (see Table S2 in the supplemental material), four strains from the Schmale collection, and 17 strains from GenBank (see Table S1 in the supplemental material). Strain numbers or GenBank accession numbers follow the underscores. *, species whose full-length TRI101/TRI201 gene products were analyzed in this study for their ability to detoxify DON. Numbers at branch points represent bootstrap values.

Article Snippet: The TRI101/TRI201 proteins were diluted to 0.2 mg/ml in a degassed buffer containing 100 mM potassium phosphate at pH 8, and spectra were recorded from 190 to 260 nm in a model 202SF circular dichroism spectrophotometer (Aviv Biomedical).

Techniques: Sequencing

Journal: Applied and Environmental Microbiology

Article Title: Bioprospecting for Trichothecene 3- O -Acetyltransferases in the Fungal Genus Fusarium Yields Functional Enzymes with Different Abilities To Modify the Mycotoxin Deoxynivalenol ▿ †

doi: 10.1128/AEM.01738-10

Figure Lengend Snippet: Pairwise comparisons between full-length TRI101 and TRI201 DNA sequences evaluated in the present study a

Article Snippet: The TRI101/TRI201 proteins were diluted to 0.2 mg/ml in a degassed buffer containing 100 mM potassium phosphate at pH 8, and spectra were recorded from 190 to 260 nm in a model 202SF circular dichroism spectrophotometer (Aviv Biomedical).

Techniques:

Journal: Applied and Environmental Microbiology

Article Title: Bioprospecting for Trichothecene 3- O -Acetyltransferases in the Fungal Genus Fusarium Yields Functional Enzymes with Different Abilities To Modify the Mycotoxin Deoxynivalenol ▿ †

doi: 10.1128/AEM.01738-10

Figure Lengend Snippet: Kinetic constant K m and k cat values and T m values for TRI101/TRI201 enzymes a

Article Snippet: The TRI101/TRI201 proteins were diluted to 0.2 mg/ml in a degassed buffer containing 100 mM potassium phosphate at pH 8, and spectra were recorded from 190 to 260 nm in a model 202SF circular dichroism spectrophotometer (Aviv Biomedical).

Techniques: Variant Assay

Journal: Applied and Environmental Microbiology

Article Title: Bioprospecting for Trichothecene 3- O -Acetyltransferases in the Fungal Genus Fusarium Yields Functional Enzymes with Different Abilities To Modify the Mycotoxin Deoxynivalenol ▿ †

doi: 10.1128/AEM.01738-10

Figure Lengend Snippet: Western blot of TRI101/TRI201 enzymes from F. pseudograminearum (Fps2TRI101), F. graminearum (Fg1TRI101), F. crookwellense (Fcr2TRI101), F. culmorum (Fcu2TRI201), F. fujikuroi (Ff1TRI201), F. sporotrichioides (Fs1TRI101), and F. oxysporum (Fo1TRI201) (1 and 2 indicate that the strain is from the Schmale or Leslie collection, respectively). Protein extracts from yeast cultures were collected following 24 h of expression. Untransformed yeast strain RW2802 represents the negative control. Ten microliters of each protein extract was loaded onto a 12% acrylamide-SDS-PAGE gel and run at 150 V for 1 h. Precision plus protein dual color standard was used to determine protein size. Ten microliters of TRI101 from F. sporotrichioides and TRI201 from F. fujikuroi, both purified from E. coli, were loaded as positive controls at known amounts of 1,000 ng, 250 ng, and 31.25 ng. The Western blot was probed with rabbit anti-FsTRI101 primary antibody and detected with alkaline phosphatase-conjugated anti-rabbit antibody.

Article Snippet: The TRI101/TRI201 proteins were diluted to 0.2 mg/ml in a degassed buffer containing 100 mM potassium phosphate at pH 8, and spectra were recorded from 190 to 260 nm in a model 202SF circular dichroism spectrophotometer (Aviv Biomedical).

Techniques: Western Blot, Expressing, Negative Control, SDS Page, Purification