Team:UC Davis/Adding secretion

 Adding Secretion               </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);"> <div style="text-align: left; background-color: rgb(255, 255, 255);"><big style="text-decoration: underline;"> <span style="font-weight: bold;"> Adding Secretion: <big style="font-weight: bold;">General model for secretion</a> Secretion Models</a> Why test different genes</a> <hr style="width: 100%; height: 2px;"><big style="text-decoration: underline;"> <span style="font-weight: bold;"> <p class="MsoNormal" style="line-height: normal;"><span style="font-size: 12pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p>  <span style="font-size: 20pt; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"> </b><big style="text-decoration: underline;"> <span style="font-weight: bold;"> <span style="font-weight: bold;"> <span style="text-decoration: underline; background-color: rgb(255, 255, 0);"></a>General model for secretion system: <img style="width: 242px; height: 58px; background-color: rgb(255, 255, 255);" alt="" src="http://2009.igem.org/wiki/images/4/4e/UCDAVIS_PROMOTER_1.png"><img style="width: 89px; height: 81px; background-color: rgb(255, 255, 255);" alt="" src="http://2009.igem.org/wiki/images/1/15/UCDAVIS_RBS1.png"><img style="width: 106px; height: 56px; background-color: rgb(255, 255, 255);" alt="" src="http://2009.igem.org/wiki/images/9/95/UCDAVIS_SECRETION.png"><img style="width: 94px; height: 69px; background-color: rgb(255, 255, 255);" alt="" src="http://2009.igem.org/wiki/images/9/9d/UCDAVIS_SS1.png"><img style="width: 106px; height: 59px; background-color: rgb(255, 255, 255);" alt="" src="http://2009.igem.org/wiki/images/0/0c/UCDAVIS_GENE1.png"><img style="width: 134px; height: 66px; background-color: rgb(255, 255, 255);" alt="" src="http://2009.igem.org/wiki/images/c/c0/UCDAVIS_6HIS.png"><img style="width: 94px; height: 83px; background-color: rgb(255, 255, 255);" alt="" src="http://2009.igem.org/wiki/images/c/ca/UCDAVIS_STOP.png"> <span style="text-decoration: underline;"> <p class="MsoNormal" style="text-align: left;"> <span style="font-family: Times New Roman,Times,serif;">The purpose of the secretion system is to introduce a method of secreting target proteins we wish to synthesize in the marvelous host: E.coli. Park <i style="font-family: Times New Roman,Times,serif;">et. al</i><span style="font-family: Times New Roman,Times,serif;"> showed that the truncated form of ice nucleation protein (INPNC) containing the complete coding region of phaZ1, including its signal sequence, could lead to stable secretion of the enzyme ( 15</a><span style="font-family: Times New Roman,Times,serif;">). We have decided to see if we could co-opt this system to make a more generalized inducible system for protein secretion in which the INPNC and phaZ1 signal sequence serve as carriers for a target protein. <o:p style="font-family: Times New Roman,Times,serif;"></o:p> <p class="MsoNormal" style="font-family: Times New Roman,Times,serif;"> <span style=""> Possible challenges: <o:p></o:p> <p class="MsoNormal" style="margin-left: 30pt; text-align: left; font-family: Times New Roman,Times,serif;"> <span style="">1. Efficient recognition of the cleavage site may require<span style="-moz-background-clip: -moz-initial; -moz-background-origin: -moz-initial; -moz-background-inline-policy: -moz-initial; background-attachment: scroll;"><span style="background: yellow none repeat scroll 0% 50%; -moz-background-clip: -moz-initial; -moz-background-origin: -moz-initial; -moz-background-inline-policy: -moz-initial;"> some of the non-signal sequence portion of the phaZ1 protein (we have only cloned the signal sequence up to the cleavage site). 2. The system may only work for proteins in a narrow site range. 3. The expression levels of the system may be low.<o:p></o:p> <p class="MsoNormal" style="text-align: center; font-family: Times New Roman,Times,serif;" align="center">  Advantages: </b> <o:p></o:p> <ol style="text-align: left;" start="1" type="1"> <li style="font-family: Times New Roman,Times,serif;" class="MsoNormal"> <u1:p></u1:p> In our secretion system, we are using two genes with different sizes as our target secretion genes; GFP being short in length and Luciferase being comparably long, these would test the ability of our secretion system to secrete proteins of different sizes. <o:p></o:p> </li> <li style="font-family: Times New Roman,Times,serif;" class="MsoNormal"> Testing various combinations of the Signal Sequence with either ompA or INPNC would help us find the best combination for our secretion system based on their secretion ability and allow us to rank the combinations from strongest to weakest in strength. (in this specific system).<o:p></o:p> </li> <li class="MsoNormal"> <big style="font-family: Times New Roman,Times,serif;"> We have submitted the BioBricks to the parts registry; therefore they can be used in future studies (for different purposes). <o:p></o:p> </li> </ol> <big style="font-family: Times New Roman,Times,serif;"> <span style="font-weight: bold;"> <span style="text-decoration: underline; background-color: rgb(255, 255, 0);"> <hr style="width: 100%; height: 2px;"> <big style="font-weight: bold;"> </a>Secretion Models: Click on a specific model for more information: <big style="font-weight: bold;"> <span style="text-decoration: underline;"> <span style="font-weight: bold;"><img alt="" src="http://2009.igem.org/wiki/images/b/ba/UCDAVIS_PIC12.png" style="border: 0px solid ; width: 95px; height: 63px;"></a><img alt="" src="http://2009.igem.org/wiki/images/a/a4/UCDAVIS_PIC13.png" style="border: 0px solid ; width: 95px; height: 63px;"></a> <span style="text-decoration: underline; font-weight: bold;"> <span style="text-decoration: underline; font-weight: bold;"> <span style="text-decoration: underline;"> <p class="MsoNormal"><span style="font-size: 13.5pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p> </b> <big style="font-weight: bold;"> <hr style="width: 100%; height: 2px;">  <span style="font-size: 20pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"> <a name="different_genes"></a>Why test different genes in our secretion system? <o:p></o:p> </b> <p class="MsoNormal"><b><span style="font-size: 13.5pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><span style="">     </b><span style="font-size: 13.5pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"> There is wide range of genes present in our environment. Therefore, it is important to measure our secretion system’s ability to secrete genes of various sizes. <b><span style="font-size: 13.5pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"> <span style="text-decoration: underline;"> </b> <b><span style="font-size: 13.5pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><span style="text-decoration: underline;">Molecular data on some proteins <o:p></o:p> </b> <p style="text-align: center;" class="MsoNormal"><b><span style="font-size: 14pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><br style=""> </b><b> <span style="font-size: 12pt; line-height: 115%; font-family: &quot;Times New Roman&quot;,&quot;serif&quot;;"><o:p></o:p> </b> <big style="text-decoration: underline;"> <span style="font-weight: bold;"> <big style="text-decoration: underline;"> <span style="font-weight: bold;">