Figure S1 - <em>In-Vitro</em> Helix Opening of <em>M. tuberculosis oriC</em> by DnaA Occurs at Precise Location and Is Inhibited by IciA Like Protein

IciA cannot inhibit helix unwinding once DnaA has already opened the DNA double helix. The reaction was carried out using 0.2 μg of DnaA protein. Helix opening was monitored by primer extension using SeqOriR1 on a 6% sequencing gel. The different lanes are: lane 3, With DnaA but without rIciA; lane 4: With DnaA and IciA (0.4 μg), but IciA was added after 10 min the addition of DnaA at 37°C. Arrows correspond to the extension products of 200 and 199 nucleotides. Lane 1 and 2 show 50 bp and 10 bp marker. (1.05 MB EPS)当DnaA（DnaA）已打开DNA双螺旋后，IciA（IciA）便无法再抑制螺旋解旋过程。本实验使用0.2 μg的DnaA蛋白完成反应体系构建。螺旋解旋情况通过以SeqOriR1为引物的引物延伸实验，在6%测序凝胶上进行监测。各组泳道设置如下：泳道3：仅添加DnaA，不添加rIciA；泳道4：同时添加DnaA与0.4 μg的IciA，但IciA是在37℃下向体系中加入DnaA 10分钟后再添加的。箭头所指为长度200和199核苷酸的延伸产物。泳道1和泳道2分别对应50 bp与10 bp分子量标记。(文件大小：1.05 MB，格式为EPS)