Difference between revisions of "Template:Virginia/Composite Part"

 
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<h1 id="composite-parts">Composite Parts</h1>
 
<h1 id="composite-parts">Composite Parts</h1>
<table style="width:100%" id="parts">
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<table style="width:100%" class="va-parts">
 
     <tr>
 
     <tr>
 
         <th>Name</th>
 
         <th>Name</th>
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     <tr>
 
     <tr>
 
         <td>LuxS-Block</td>
 
         <td>LuxS-Block</td>
         <td>(spacerLXf (K2535003) + I719005 + B0034 + K2535000 + B0015 + spacerLXr (K2535004)). LuxS is a protein, not native to the Lsr Operon, that catalyzes the cleavage of metabolites to synthesize the autoinducer molecule AI-2 (Gonzalez et al 2006). The LuxS gene is regulated by a T7 driven promoter in order to cooperate with the AI-2 sensitive plasmid that this part is used in conjunction with.  
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         <td>spacer_pT7_rbs_LuxS_term_spacer. LuxS is a protein, not native to the Lsr Operon, that catalyzes the cleavage of metabolites to synthesize the autoinducer molecule AI-2 (Gonzalez et al 2006). The LuxS gene is regulated by a T7 driven promoter in order to cooperate with the AI-2 sensitive plasmid that this part is used in conjunction with. </td>
        </td>
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         <td> K2535005 </td>
         <td> spacer_pT7_rbs_LuxS_term_spacer -- BBa_K2535005 <em>SENT AS A BIOBRICK IN pSB1C3</em></td>
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     </tr>
 
     </tr>
 
     <tr>
 
     <tr>
         <td>LuxS 2nd Plasmid (LuxS-sfGFP)</td>
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         <td>LuxS-sfGFP</td>
         <td>I746909 + K2535005. This is a composite part of LuxS-Block and I746909. This part can be regulated using T7 to track LuxS expression based on the sfGFP reporter. Both LuxS and sfGFP are controlled by a T7 promoter. </td>
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         <td> pT7_rbs_sfGFP_term_LuxS-Block. This is a composite part of LuxS-Block and I746909. This part can be regulated using T7 to track LuxS expression based on the sfGFP reporter. Both LuxS and sfGFP are controlled by a T7 promoter.</td>
         <td> pT7_rbs_sfGFP_term_LuxS-Block  -- K2535006</td>
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         <td> K2535006</td>
 
     </tr>
 
     </tr>
 
     <tr>
 
     <tr>
 
         <td>LsrK-Block</td>
 
         <td>LsrK-Block</td>
         <td>spacerLRf(K2535007) + I719005 + B0034 + K091002 + B0015 + spacerLRr (K2535008). LsrK is a kinase that is native to the Lsr Operon. It converts intracellular AI-2 to phosphorylated AI-2 (AI-2P), which derepresses the Lsr Operon (Xavier et al 2004). The LsrK gene is regulated by a T7 driven promoter in order to cooperate with the AI-2 sensitive plasmid that this part is used in conjunction with. </td>
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         <td>spacer_ pT7_rbs_LsrK_term_spacer. LsrK is a kinase that is native to the Lsr Operon. It converts intracellular AI-2 to phosphorylated AI-2 (AI-2P), which derepresses the Lsr Operon (Xavier et al 2004). The LsrK gene is regulated by a T7 driven promoter in order to cooperate with the AI-2 sensitive plasmid that this part is used in conjunction with. </td>
         <td> spacer_ pT7_rbs_LsrK_term_spacer -- K2535009</td>
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         <td>K2535009</td>
 
     </tr>
 
     </tr>
 
     <tr>
 
     <tr>
         <td>LsrK 2nd Plasmid (LsrK-sfGFP)</td>
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         <td>LsrK-sfGFP</td>
         <td>I746909 + K2535009. This is a composite part of LsrK-Block and I746909. This part can be regulated using T7 to track LsrK expression based on the sfGFP reporter. Both LsrK and sfGFP are controlled by a T7 promoter. </td>
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         <td>pT7_rbs_sfGFP_term_LsrK-Block. This is a composite part of LsrK-Block and I746909. This part can be regulated using T7 to track LsrK expression based on the sfGFP reporter. Both LsrK and sfGFP are controlled by a T7 promoter.</td>
         <td>pT7_rbs_sfGFP_term_LsrK-Block -- K2535010</td>
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         <td>K2535010</td>
 
     </tr>
 
     </tr>
 
     <tr>
 
     <tr>
 
         <td>YdgG-Block</td>
 
         <td>YdgG-Block</td>
         <td>spacerYf(K2535011) + I719005 + B0034 + K2535002 + B0015 + spacerYr(K2535012).YdgG is a membrane transport protein, not native to the Lsr Operon, which exports AI-2 out of the cell (Herzberg et al 2006). The YdgG gene is regulated by a T7 driven promoter in order to cooperate with the AI-2 sensitive plasmid that this part is used in conjunction with. </td>
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         <td>spacer_ pT7_rbs_YdgG_term_spacer. YdgG is a membrane transport protein, not native to the Lsr Operon, which exports AI-2 out of the cell (Herzberg et al 2006). The YdgG gene is regulated by a T7 driven promoter in order to cooperate with the AI-2 sensitive plasmid that this part is used in conjunction with. </td>
         <td>spacer_ pT7_rbs_YdgG_term_spacer -- K2535013</td>
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         <td>K2535013</td>
 
     </tr>
 
     </tr>
 
     <tr>
 
     <tr>
         <td>YdgG 2nd Plasmid (YdgG-sfGFP)</td>
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         <td>YdgG-sfGFP</td>
         <td>I746909 + K2535013. This is a composite part of YdgG-Block and I746909. This part can be regulated using T7 to track YdgG expression based on the sfGFP reporter. Both YdgG and sfGFP are controlled by a T7 promoter. </td>
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         <td>pT7_rbs_sfGFP_term_YdgG-Block. This is a composite part of YdgG-Block and I746909. This part can be regulated using T7 to track YdgG expression based on the sfGFP reporter. Both YdgG and sfGFP are controlled by a T7 promoter. </td>
         <td>pT7_rbs_sfGFP_term_YdgG-Block -- K2535014</td>
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         <td>K2535014</td>
 
     </tr>
 
     </tr>
 
     <tr>
 
     <tr>
         <td>LsrK-LuxS 2nd Plasmid (LsrK-LuxS-sfGFP)</td>
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         <td>LsrK-LuxS-sfGFP</td>
         <td>I746909 + K2535009 + K2535005. This is a composite part of LsrK and LuxS put together with I746909 (sfGFP regulated by a T7 promoter). This part allows the upregulation of LuxS and LsrK so that intracellular AI-2 production and phosphorylation are increased concurrently. </td>
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         <td>pt7_rbs_sfGFP_term_LsrK-Block_LuxS-Block. This is a composite part of LsrK and LuxS put together with I746909 (sfGFP regulated by a T7 promoter). This part allows the upregulation of LuxS and LsrK so that intracellular AI-2 production and AI-2 phosphorylation are increased concurrently. </td>
         <td> pt7_rbs_sfGFP_term_LsrK-Block_LuxS-Block -- K2535015</td>
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         <td> K2535015</td>
 
     </tr>
 
     </tr>
 
     <tr>
 
     <tr>
         <td>LsrK-LuxS-YdgG 2nd Plasmid (LsrK-LuxS-YdgG-sfGFP)</td>
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         <td>LsrK-YdgG-sfGFP</td>
         <td>I746909 + K2535009 + K2535005 + K2535013. This is a composite part of LsrK, LuxS, and YdgG put together with I746909 (sfGFP regulated by a T7 promoter).  This part allows the upregulation of LsrK, LuxS, and YdgG so that intracellular AI-2 production, AI-2 phosphorylation, and export out of the cell are all increased concurrently. </td>
+
         <td>This is a composite part of LsrK and YdgG put together with I746909 (sfGFP regulated by a T7 promoter).  This part allows the upregulation of LsrK and YdgG so that AI-2 is phosphorylated to AI-2P and AI-2 out of the cell are increased concurrently. </td>
         <td> pt7_rbs_sfGFP_term_LsrK_LuxS_YdgG -- K2535017 <em>SENT AS A BIOBRICK IN pSB1C3</em> </td>
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         <td>K2535016 </td>
 
     </tr>
 
     </tr>
 
     <tr>
 
     <tr>
         <td>sQS: Synthetic Quorum Response Plasmid Insert (1st Plasmid)</td>
+
        <td>LuxS-YdgG-sfGFP</td>
         <td>B0014 (term) + K2535018 + K145001 (T7) + B0015 (term). Built in low copy pACYC cloning vector and optimized for use in a low copy vector to prevent leakiness of AI-2 regulated promoter. This plasmid contains an AI-2 regulated promoter, pLsr, which is bidirectional. Dimers of LsrR bind to and repress this promoter. However, AI-2P can bind to these dimers and cause them to dissociate, depressing the promoter.  Since the expression of this plasmid produces LsrR, which represses the promoter pLsr, this system is self regulating. sQS plasmid is used to regulate quorum enhancement plasmids. T7 is therefore only expressed in the presence of AI-2P. This plasmid can be used to regulate T7 expression based on AI-2 level in a media.  
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        <td>pt7_rbs_sfGFP_term_LuxS-Block_YdgG-Block. This is a composite part of LuxS and YdgG put together with I746909 (sfGFP regulated by a T7 promoter).  This part allows the upregulation of LuxS and YdgG so that intracellular AI-2 production and export out of the cell are increased concurrently.</td>
        </td>
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        <td>K2535020</td>
         <td>term_pLsr_rbs_LsrR_rbs_T7_term -- K2535019</td>
+
    </tr>
 +
    <tr>
 +
        <td>LsrK-LuxS-YdgG-sfGFP</td>
 +
        <td>pt7_rbs_sfGFP_term_LsrK_LuxS_YdgG. This is a composite part of LsrK, LuxS, and YdgG put together with I746909 (sfGFP regulated by a T7 promoter).  This part allows the upregulation of LsrK, LuxS, and YdgG so that intracellular AI-2 production, AI-2 phosphorylation, and export out of the cell are all increased concurrently.  </td>
 +
        <td> K2535017 </td>
 +
    </tr>
 +
    <tr>
 +
         <td>sQS: Synthetic Quorum Response Plasmid Insert</td>
 +
         <td>term_pLsr_rbs_LsrR_rbs_T7_term. Built in low copy pACYC cloning vector and optimized for use in a low copy vector to prevent leakiness of AI-2 regulated promoter. This plasmid contains an AI-2 regulated promoter, pLsr, which is bidirectional. Dimers of LsrR bind to and repress this promoter. However, AI-2P can bind to these dimers and cause them to dissociate, depressing the promoter.  Since the expression of this plasmid produces LsrR, which represses the promoter pLsr, this system is self regulating. sQS plasmid is used to regulate quorum enhancement plasmids. T7 is therefore only expressed in the presence of AI-2P. This plasmid can be used to regulate T7 expression based on AI-2 level in a media. </td>
 +
         <td>K2535019</td>
 
     </tr>
 
     </tr>
 
</table>
 
</table>
 +
 +
<h1 id="references">References</h1>
 +
<p>Gonzalez J, Neshavan N (2006). Messing with Bacterial Quorum Sensing. Microbiology and Molecular Biology
 +
Reviews 70, 859-875.    </p>
 +
<p>Herzberg M, Kaye I, Peti W, Wood T (2006) YdgG (TqsA) Controls Biofilm Formation in Escherichia
 +
coli K-12 through Autoinducer 2 Transport. J Bacteriol. 2006 Jan; 188(2): 587–598.        </p>
 +
<p>Xavier K, Bassler B (2004). Regulation of Uptake and Processing of the Quorum-Sensing Autoinducer AI-2 in
 +
Escherichia coli. J Bacteriol 187, 238-245.      </p>

Latest revision as of 20:12, 17 October 2018

Composite Parts

Name Description Part
LuxS-Block spacer_pT7_rbs_LuxS_term_spacer. LuxS is a protein, not native to the Lsr Operon, that catalyzes the cleavage of metabolites to synthesize the autoinducer molecule AI-2 (Gonzalez et al 2006). The LuxS gene is regulated by a T7 driven promoter in order to cooperate with the AI-2 sensitive plasmid that this part is used in conjunction with. K2535005
LuxS-sfGFP pT7_rbs_sfGFP_term_LuxS-Block. This is a composite part of LuxS-Block and I746909. This part can be regulated using T7 to track LuxS expression based on the sfGFP reporter. Both LuxS and sfGFP are controlled by a T7 promoter. K2535006
LsrK-Block spacer_ pT7_rbs_LsrK_term_spacer. LsrK is a kinase that is native to the Lsr Operon. It converts intracellular AI-2 to phosphorylated AI-2 (AI-2P), which derepresses the Lsr Operon (Xavier et al 2004). The LsrK gene is regulated by a T7 driven promoter in order to cooperate with the AI-2 sensitive plasmid that this part is used in conjunction with. K2535009
LsrK-sfGFP pT7_rbs_sfGFP_term_LsrK-Block. This is a composite part of LsrK-Block and I746909. This part can be regulated using T7 to track LsrK expression based on the sfGFP reporter. Both LsrK and sfGFP are controlled by a T7 promoter. K2535010
YdgG-Block spacer_ pT7_rbs_YdgG_term_spacer. YdgG is a membrane transport protein, not native to the Lsr Operon, which exports AI-2 out of the cell (Herzberg et al 2006). The YdgG gene is regulated by a T7 driven promoter in order to cooperate with the AI-2 sensitive plasmid that this part is used in conjunction with. K2535013
YdgG-sfGFP pT7_rbs_sfGFP_term_YdgG-Block. This is a composite part of YdgG-Block and I746909. This part can be regulated using T7 to track YdgG expression based on the sfGFP reporter. Both YdgG and sfGFP are controlled by a T7 promoter. K2535014
LsrK-LuxS-sfGFP pt7_rbs_sfGFP_term_LsrK-Block_LuxS-Block. This is a composite part of LsrK and LuxS put together with I746909 (sfGFP regulated by a T7 promoter). This part allows the upregulation of LuxS and LsrK so that intracellular AI-2 production and AI-2 phosphorylation are increased concurrently. K2535015
LsrK-YdgG-sfGFP This is a composite part of LsrK and YdgG put together with I746909 (sfGFP regulated by a T7 promoter). This part allows the upregulation of LsrK and YdgG so that AI-2 is phosphorylated to AI-2P and AI-2 out of the cell are increased concurrently. K2535016
LuxS-YdgG-sfGFP pt7_rbs_sfGFP_term_LuxS-Block_YdgG-Block. This is a composite part of LuxS and YdgG put together with I746909 (sfGFP regulated by a T7 promoter). This part allows the upregulation of LuxS and YdgG so that intracellular AI-2 production and export out of the cell are increased concurrently. K2535020
LsrK-LuxS-YdgG-sfGFP pt7_rbs_sfGFP_term_LsrK_LuxS_YdgG. This is a composite part of LsrK, LuxS, and YdgG put together with I746909 (sfGFP regulated by a T7 promoter). This part allows the upregulation of LsrK, LuxS, and YdgG so that intracellular AI-2 production, AI-2 phosphorylation, and export out of the cell are all increased concurrently. K2535017
sQS: Synthetic Quorum Response Plasmid Insert term_pLsr_rbs_LsrR_rbs_T7_term. Built in low copy pACYC cloning vector and optimized for use in a low copy vector to prevent leakiness of AI-2 regulated promoter. This plasmid contains an AI-2 regulated promoter, pLsr, which is bidirectional. Dimers of LsrR bind to and repress this promoter. However, AI-2P can bind to these dimers and cause them to dissociate, depressing the promoter. Since the expression of this plasmid produces LsrR, which represses the promoter pLsr, this system is self regulating. sQS plasmid is used to regulate quorum enhancement plasmids. T7 is therefore only expressed in the presence of AI-2P. This plasmid can be used to regulate T7 expression based on AI-2 level in a media. K2535019

References

Gonzalez J, Neshavan N (2006). Messing with Bacterial Quorum Sensing. Microbiology and Molecular Biology Reviews 70, 859-875.

Herzberg M, Kaye I, Peti W, Wood T (2006) YdgG (TqsA) Controls Biofilm Formation in Escherichia coli K-12 through Autoinducer 2 Transport. J Bacteriol. 2006 Jan; 188(2): 587–598.

Xavier K, Bassler B (2004). Regulation of Uptake and Processing of the Quorum-Sensing Autoinducer AI-2 in Escherichia coli. J Bacteriol 187, 238-245.