<p>(A) Purified YhaJ was tested for its ability to bind the <i>yhaO</i> promoter region (<i>pyhaO;</i> ~300 bp region upstream of the <i>yhaO</i> coding sequence) by EMSA. DIG-labeled <i>pyhaO</i> was incubated with increasing concentrations of YhaJ that corresponded to a shift in free-DNA indicating a YhaJ-DNA complex. Specificity of the binding reaction was tested by the addition of a 100-fold excess (+) of unlabeled <i>pyhaO</i> probe to the binding reaction to outcompete binding of the DIG-labeled probe to YhaJ. These reaction conditions were carried out using a fragment of the <i>kan</i> gene as a negative control. Additionally, the unlabeled <i>kan</i> probe in 100-fold excess was used as a non-specific competitor for YhaJ binding to the DIG-labeled <i>yhaO</i> probe (p<i>yhaO</i> vs <i>kan</i>). (B) Activity of the <i>yhaO</i> promoter in the <i>ΔyhaJ</i> mutant background. A plasmid containing a GFP-<i>yhaO</i> promoter fusion was transformed into TUV93-0 and <i>ΔyhaJ</i> to monitor transcription of <i>yhaO</i> in RFU during growth in MEM-HEPES. Data was calculated from three biological replicates and plotted at increasing OD<sup>600</sup> values. * denotes P ≤ 0.05.</p

Similar works

Full text

oai:figshare.com:article/1632511Last time updated on 2/12/2018

This paper was published in FigShare.

Having an issue?

Is data on this page outdated, violates copyrights or anything else? Report the problem now and we will take corresponding actions after reviewing your request.