The oligonucleotides used in this study.

<p>The oligonucleotides used in this study.</p

Genomic organization suggest a LTTR-dependent regulation of <i>lysE</i>(^Mt).

<p>A 108 bp region separates <i>lysG</i>(^Mt) from <i>lysE</i>(^Mt). The <i>lysG</i>(^Mt) and <i>lysE</i>(^Mt) transcriptional start sites are indicated with +1 (in red and bold type). The coloured arrows denote the beginning of the ORFs. The promoter binding motifs (-10 and -35 motifs) are marked as boxes. The underlined base pairs represent a palindromic DNA sequence.</p

Log-log plot of the whole transcriptome expression analysis.

<p>The log-log plot shows the signal intensities of the mutant strain Δ<i>lysG</i> plotted against those of the Wt <i>Mt</i> strain. Spots represent the average of log2 data for each gene from three independent experiments. High through low expression levels are presented as a three-color spectrum. The red color indicates high expression and the green color low expression.</p

Sequence accession number received from the manufacturer.

<p>Sequence accession number received from the manufacturer.</p

Genes regulated by LysG(^Mt).

<p>Genes regulated by LysG(^Mt).</p

LysG(^Mt) upregulates <i>lysE</i>(^Mt) in the presence of lysine and histidine.

<p>Relative gene expression level of <i>lysE</i>(^Mt) in Wt <i>Mt</i> H37Rv, the knockout mutant Δ<i>lysG</i> and the complemented strain Δ<i>lysG</i>::<i>lysG</i> during incubation in a minimal medium with lysine, histidine, arginine, asparagine, aspartate and leucine as additional nitrogen source. For normalization, the expression level of the validated reference gene <i>sigA</i> and <i>sigE</i> were used. The cutoff for significant regulation was set at 2-fold change. Means were calculated from three independent experiments. Error bars represent standard errors of the mean.</p

Transcriptome profiles of the DNA containing <i>lysG</i>(^Mt) and <i>lysE</i>(^Mt).

<p>(A) Genomic region with coding sequences. (B) and (C) Profiles of mapped reads from RNAseq using RNA samples from Wt <i>Mt</i> with ammonium (B) or lysine (C) as nitrogen source. (D) and (E) Profiles of mapped reads from RNAseq using RNA samples from the Δ<i>lysG</i> mutant strain with ammonium (D) or lysine (E) as nitrogen source.</p

LysG(^Mt) binds to the upstream region of <i>lysE</i>(^Mt).

<p>(A) Intergenic region between <i>lysG</i>(^Mt) and <i>lysE</i>(^Mt). The <i>lysG</i>(^Mt) and <i>lysE</i>(^Mt) transcriptional start sites are indicated with +1 (in red and bold type). The promoter binding motifs (-10 and -35 motifs) are marked as boxes. The coloured arrows denote the beginning of the open reading frames. The location of the inverted repeat within the intergenic region is marked (underlined). The position of the labeled oligonucleotides used for the electrophoretic mobility shift assay is shown (I to IV). 30 fmole of labeled oligonucleotide I was incubated with increasing concentrations of LysG(^Mt), indicated for each lane. As control no protein (lane 1) and 50, 100 or 200-fold molar excess of unlabeled target DNA (oligonucleotide I) were added, marked in red. 5 mM of the co-effector lysine (lane 14) and 3.34 mM of histidine were added (lane 15). Bands corresponding to free DNA are marked by open arrowheads, whereas bands with DNA in complex with LysG(^Mt) are marked by filled arrowheads. Results are representative of two independent experiments. (B) Electrophoretic mobility shift assay with 200 pmole of LysG(^Mt) was done in combination with 10 fmole of the three subfragments (oligonucleotides II to IV). As control no protein (lane 4) and 200-fold molar excess of unlabeled target DNA (lane 5) were added.</p

Transcriptome profiles of the DNA region from <i>fadD26</i> to <i>ppsE</i>.

<p>(A) Genomic organization of the genes <i>fadD26</i> to <i>ppsE</i> of <i>Mt</i> is shown. (B) and (C) Profiles of mapped reads from RNAseq using RNA samples from Wt <i>Mt</i> with ammonium (B) or lysine (C) as nitrogen source. (D) and (E) Profiles of mapped reads from RNAseq using RNA samples from the Δ<i>lysG</i> mutant strain with ammonium (D) or lysine (E) as nitrogen source.</p

On Dropping Nuclear Weapons: A Question for the Ages

The United States’ decision to drop nuclear bombs on Japan remains a controversial decision to this day. The loss of life and overall destruction was so great that no country has used nuclear bombs since. However, in considering the situation in which the bombs were dropped -- warfare in the Pacific was particularly brutal -- the decision is fully justifiable. By examining great thinkers and philosophies from 1582-1988, we can understand the motivations behind wars and how best to end them. I argue that throughout the modern age, philosophers have argued that choosing the lesser of two evils is a moral responsibility. I would further assert that, given the alternatives, bombing Hiroshima and Nagasaki was definitely the lesser of two evils