Team:UC Davis/PhoR ChvI1

 PhoR to ChvI               <b style="color: rgb(255, 255, 153);"><span style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"> </b><b style="color: rgb(255, 255, 153);"><span style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"> </b><b style="color: rgb(255, 255, 153);"><span style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"> </b><b style="color: rgb(255, 255, 153);"><span style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">   <img alt="" src="http://2009.igem.org/wiki/images/b/b9/UCDAVIS_PIC8.png" style="border: 0px solid ; width: 78px; height: 36px;"></a>   </b><b style="color: rgb(255, 255, 153);"><span style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">   <img alt="" src="http://2009.igem.org/wiki/images/2/2f/UCDAVIS_PIC5.png" style="border: 0px solid ; width: 81px; height: 36px;"></a>   </b><b style="color: rgb(255, 255, 153);"><span style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"> </b><b style="color: rgb(255, 255, 153);"><span style="font-size: 13.5pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;">   <img alt="" src="http://2009.igem.org/wiki/images/a/a6/UCDAVIS_PIC6.png" style="border: 0px solid ; width: 78px; height: 37px;"></a> <img alt="" src="http://2009.igem.org/wiki/images/1/1d/UCDAVIS_PIC7.png" style="border: 0px solid ; width: 83px; height: 37px;"></a>   </b> <hr style="width: 100%; height: 2px; background-color: rgb(255, 255, 255);"> <p class="MsoNormal" style="background: white none repeat scroll 0% 50%; line-height: normal; -moz-background-clip: -moz-initial; -moz-background-origin: -moz-initial; -moz-background-inline-policy: -moz-initial;"><span style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;; color: black;"> <span style="text-decoration: underline;"> PhoR--&gt;ChvI </b><span style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;; color: black;">It has been hypothesized that due to the amino acid similarity of ChvI to PhoB, ChvI is able to act as a phosphate regulator in E.coli as a PhoB substitute (3). <span style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;; color: black;"><o:p></o:p> <p class="MsoNormal" style="background: white none repeat scroll 0% 50%; line-height: normal; -moz-background-clip: -moz-initial; -moz-background-origin: -moz-initial; -moz-background-inline-policy: -moz-initial;"><span style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;; color: black;"> Since ChvI showed no abnormal activity inside Agrobacterium tumefaciens under phosphate limitation, it cannot serve as the primary phosphate regulator in A. tumefaciens (3).<o:p></o:p> <p class="MsoNormal" style="background: white none repeat scroll 0% 50%; line-height: normal; -moz-background-clip: -moz-initial; -moz-background-origin: -moz-initial; -moz-background-inline-policy: -moz-initial;"><span style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;; color: black;"><span style=""> <i>Note: ChvI was not required for phosphate regulation in A. tumefaciens (3), but it might be involved in phosphate regulation in A. tumefaciens in some way (3). Therefore, the possibility of crosstalk still exists</i><o:p></o:p> <p class="MsoNormal" style="background: white none repeat scroll 0% 50%; line-height: normal; -moz-background-clip: -moz-initial; -moz-background-origin: -moz-initial; -moz-background-inline-policy: -moz-initial;"><span style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;; color: black;"> <span style="">  Due to the structural similarity of ChvI and PhoB, the possibility of crosstalk exists (3).<o:p></o:p> <p class="MsoNormal"><i><span style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;; color: black;"> Note: Since we are going to use E. coli as our model chassis for this project; we had to make sure that our proteins will keep their characteristics of our interest after being placed inside the E. coli. </i><span style="font-size: 13pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;; color: black;"> <i>For example, VirG gene (promoter 1) after being introduced into E. coli cells, started to behave as if it were a member of phosphate regulon (note: PhoB function was necessary for virG induction by phosphate starvation)(6).</i> <i>Studies have suggested that the reason for this behavior was due to the fact that virG promoter 1 has two hexamer blocks which have the same positions as the pho genes (6).</i> <o:p></o:p> <p class="MsoNormal" style="background: white none repeat scroll 0% 50%; line-height: normal; -moz-background-clip: -moz-initial; -moz-background-origin: -moz-initial; -moz-background-inline-policy: -moz-initial;"><span style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;; color: black;"><span style=""> Studies have shown that ChvI probably activated Alkaline Phosphatase <span style="font-size: 13pt; font-family: &quot;Calibri&quot;,&quot;sans-serif&quot;;"> <span style="font-size: 13pt;">(AP) <span style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;; color: black;"> activity by activating transcription of PhoA (PhoA also gets activated by PhoB) (3). It was not proven that ChvI activates other PhoB-dependent promoters or recognizes the same DNA sequence as PhoB (3). However, there has been speculation that since PhoR mediates PhoB phosphorylation and dephosphorylation, it may do the same to ChvI(3).<o:p></o:p> <p class="MsoNormal" style="background: white none repeat scroll 0% 50%; line-height: normal; -moz-background-clip: -moz-initial; -moz-background-origin: -moz-initial; -moz-background-inline-policy: -moz-initial;"><span style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;; color: black;"> Therefore, the best option would be to find a promoter from A. tumefaciens that gets activated by ChvG/ChvI, a two-component regulatory system only; and does not interact with any E. coli system. <span style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;; color: black;"> </b> <hr style="width: 100%; height: 2px;"> <p class="MsoNormal" style="background: white none repeat scroll 0% 50%; line-height: normal; -moz-background-clip: -moz-initial; -moz-background-origin: -moz-initial; -moz-background-inline-policy: -moz-initial;"><span style="font-size: 13pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;; color: black;"><o:p></o:p> </b>