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The minimum folding energy (MFE) of 60 nt RNA segments immediately upstream of the initiation codons was calculated as a proxy of secondary structure stability.
The minimum folding energy (MFE) was computed using DAMBE which incorporates the function library of the Vienna RNA package [22], at 37°C, with no lonely pairs and with no GU pairs at the end of helices.
We found that the IRES activity of yeast IRESs is strongly associated with weak secondary structure measured by the minimum folding energy (MFE, in kcal/mol) of the 60 nt immediately upstream of the initiation AUG (Figure 1, r = −0.7756, p = 0.0002), contrary to the conventional belief that IRESs should have complex and stable secondary structure [4] [7].
L3utr length of 3′ UTR, dG minimum folding energy.
The minimum folding energy (MFE) of the obtained 2562/1449/2475 hairpin precursors was evaluated with the UNAFold program.
The miRNAs were first filtered for the thermodynamic stability of heteroduplex between miRNA and mRNA target site, based on minimum folding energy (MFE).
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The minimum folding energies (ΔGfolding) for the first 40 nucleotides from each of the 4288 E. coli ORFs were calculated using UNAFold [ 38] with the default setting of 37°C.
We employed sequence composition-based features, the minimum folding energies (MFEs) of the RNAs, and certain orthologous group comparative properties to generate a total of 29 features, which are listed in Additional file 1 (see Methods).
The predicted minimum fold energy of 5'UTR sequences was determined using the Vienna RNAfold WebServer [ 42].
Plant sp, NSs, Nucleotide substitutions between known plant query miRNAs and the corresponding miRNA in Boechera apomictic species; NN, Number of nucleotides hairpin length; ARM, mature miRNA location in hairpin structure; AMFE, Adjusted minimum fold energy; MFEI, Minimum fold energy index; EST ID, Identifier of the 454 transcripts from which miRNA was derived.
Plant sp, NSs, Nucleotide substitutions between known plant query miRNAs and the corresponding miRNA in Boechera sexual species; NN, Number of nucleotides hairpin length; ARM, mature miRNA location in hairpin structure; AMFE, Adjusted minimum fold energy; MFEI, Minimum fold energy index; EST ID, Identifier of the 454 transcripts from which miRNA was derived.
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