Difference between revisions of "Team:JiangnanU China/Parts"

 
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             <div class="centers">
 
             <div class="centers">
 
                 <div class="fb_72">
 
                 <div class="fb_72">
−
                     <b>Part</b>
+
                     <b>Parts</b>
 
                 </div>
 
                 </div>
 
                 <div style="height: 60vh"></div>
 
                 <div style="height: 60vh"></div>
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         <div class="centers">
 
         <div class="centers">
 
             <div class="fb_72">
 
             <div class="fb_72">
−
                 <b>Basic Part</b>
+
                 <b>Basic Parts</b>
 
             </div>
 
             </div>
 
             <!--            BBa_K3137000-->
 
             <!--            BBa_K3137000-->
 
             <div class="split_small"></div>
 
             <div class="split_small"></div>
 
             <div class="fb_48">
 
             <div class="fb_48">
−
                 <b>BBa_K3137000</b>
+
                 <b>BBa_K3137000 : <a href="http://parts.igem.org/Part:BBa_K3137000" style="color:blue;"alt="">P<i>rsmH</i></a></b>
 
             </div>
 
             </div>
 
             <br/>
 
             <br/>
 
             <div class="fm_22">
 
             <div class="fm_22">
−
                 <i>rsmH</i>      50bp constitutive promoter<br/>
+
                 250bp constitutive promoter<br/>
−
                 <i>RsmH</i> can detect the intensity of the fluorescence of <i>gfp</i>.
+
                 We ligated promoter <i>rsmH</i> with <i>gfp</i> to measure the fluorescence intensity of GFP.
 
             </div>
 
             </div>
−
            <!--            BBa_K3137002-->
+
          <!--            BBa_K3137001-->
 
             <div class="split_small"></div>
 
             <div class="split_small"></div>
 
             <div class="fb_48">
 
             <div class="fb_48">
−
                 <b>BBa_K3137002</b>
+
                 <b>BBa_K3137001 : <a href="http://parts.igem.org/Part:BBa_K3137001" style="color:blue;"alt="">Improved RBS 4</a></b>
 
             </div>
 
             </div>
 
             <br/>
 
             <br/>
 
             <div class="fm_22">
 
             <div class="fm_22">
−
                 P<i>putA</i>       50bp inducible promoter<br/>
+
                 11bp RBS<br/>
 +
                As we all know, RBS refers to a sputum-rich untranslated region upstream of the initiation codon AUG.
 +
                <a href="http://parts.igem.org/wiki/index.php/Part:BBa_B0033" style="color:blue;"alt="">BBa_B0033</a> is a weaker RBS based on Ron Weiss thesis.
 +
                Compared with <a href="http://parts.igem.org/wiki/index.php/Part:BBa_B0030" style="color:blue;"alt="">BBa_B0030</a>,
 +
              <a href="http://parts.igem.org/wiki/index.php/Part:BBa_B0031" style="color:blue;"alt="">BBa_B0031</a>,
 +
              <a href="http://parts.igem.org/wiki/index.php/Part:BBa_B0032" style="color:blue;"alt="">BBa_B0032</a>,
 +
              <a href="http://parts.igem.org/wiki/index.php/Part:BBa_B0034" style="color:blue;"alt="">BBa_B0034</a>,
 +
              BBa_B0033 is the weakest,
 +
                and whose RBS strength is 0.35% of BBa_B0034. By analyzing their sequences, we found that BBa_B0033 has fewer purines and no not have AAA sequence.
 +
                SO we designed the RBS by adjusting the proportion of bases.
 +
            <br/>
 +
              Then we ligated the two RBS with promotor <i>rsmH</i> (<a href="http://parts.igem.org/wiki/index.php/Part:BBa_K3137000" style="color:blue;"alt="">BBa_K3137000</a>)
 +
            and gene <i>gfp</i> (<a href="http://parts.igem.org/wiki/index.php/Part:BBa_K3137011" style="color:blue;"alt="">BBa_K3137011</a>) and transform into <i>E. coli</i> BL21 to
 +
              compare their green fluorescence. And take <i>E. coli</i> BL21 as a control.
 +
                The figure below shows that <a href="http://parts.igem.org/wiki/index.php/Part:BBa_K3137001" style="color:blue;"alt="">BBa_K3137001</a>
 +
                has the stronger fluorescently
 +
              protein expression than BBa_B0033, that means it’s efficient to increase the protein expression by increasing the number of purines in the RBS sequence,
 +
              which is beneficial to the binding of ribosomes.
 +
            </div>
 +
        <!--            BBa_K3137002-->
 +
            <div class="split_small"></div>
 +
            <div class="fb_48">
 +
                <b>BBa_K3137002 : <a href="http://parts.igem.org/Part:BBa_K3137002" style="color:blue;"alt="">P<i>putA</i></a></b>
 +
            </div>
 +
            <br/>
 +
            <div class="fm_22">
 +
                250bp inducible promoter<br/>
 
                 P<i>putA</i> is an inducible promoter. When <i>E.coli</i> is infected by a phage, both report and
 
                 P<i>putA</i> is an inducible promoter. When <i>E.coli</i> is infected by a phage, both report and
−
                 respond gene
+
                 response gene
 
                 circuits are induced to express at the same time. When a phage infects <i>Escherichia coli</i> into the
 
                 circuits are induced to express at the same time. When a phage infects <i>Escherichia coli</i> into the
−
                incubation period (about 5 min), P<i>putA</i> will induce the bacteria to express anti-protein and
+
              latent period (about 5 min), P<i>putA</i> will induce the bacteria to express anti-protein and
 
                 display
 
                 display
 
                 green fluorescence.
 
                 green fluorescence.
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             <div class="split_small"></div>
 
             <div class="split_small"></div>
 
             <div class="fb_48">
 
             <div class="fb_48">
−
                 <b>BBa_K3137003</b>
+
                 <b>BBa_K3137003 : <a href="http://parts.igem.org/Part:BBa_K3137003" style="color:blue;"alt=""> P<i>glcF </i> </a></b></b>
 
             </div>
 
             </div>
 
             <br/>
 
             <br/>
 
             <div class="fm_22">
 
             <div class="fm_22">
−
                 P<i>glcF </i>      50bp inducible promoter<br/>
+
                 250bp inducible promoter<br/>
 
                 P<i>glcF</i> is an inducible promoter that works when phages infect bacterial for around 20 min. If the
 
                 P<i>glcF</i> is an inducible promoter that works when phages infect bacterial for around 20 min. If the
 
                 anti-proteins successfully resist phage infection, the inducible promoter P<i>glcF</i> will not
 
                 anti-proteins successfully resist phage infection, the inducible promoter P<i>glcF</i> will not
 
                 induce the
 
                 induce the
 
                 expression of downstream gene. If the anti-proteins cannot kill phages, the phages will continue to
 
                 expression of downstream gene. If the anti-proteins cannot kill phages, the phages will continue to
−
                 infect. When phages infect bacteria into the outbreak period (about 20 min), the inducible promoter
+
                 infect. When phages infect bacteria into the burst period (about 20 min), the inducible promoter
−
                 P<i>glcF</i> will induce expression of the red fluorescence protein gene <i>mCherry</i> and the
+
                 P<i>glcF</i> will induce expression of the red fluorescent protein gene <i>mCherry</i> and the
 
                 downstream toxic
 
                 downstream toxic
 
                 protein gene <i>protegrin-1</i>, which will make the bacteria display red fluorescence and lyse cells
 
                 protein gene <i>protegrin-1</i>, which will make the bacteria display red fluorescence and lyse cells
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             <div class="split_small"></div>
 
             <div class="split_small"></div>
 
             <div class="fb_48">
 
             <div class="fb_48">
−
                 <b>BBa_K3137004</b>
+
                 <b>BBa_K3137004 : <a href="http://parts.igem.org/Part:BBa_K3137004" style="color:blue;"alt=""><i>mCherry</i></a></b>
 
             </div>
 
             </div>
 
             <br/>
 
             <br/>
 
             <div class="fm_22">
 
             <div class="fm_22">
−
                 <i>mCherry</i>      708bp report gene<br/>
+
                 708bp reporter gene<br/>
−
                 The <i>mCherry</i> can express red fluorescence protein and it is used as a report  gene in the process of
+
                 The <i>mCherry</i> can express red fluorescent protein and it is used as a reporter gene in the process of
 
                 reporting the number of cells.
 
                 reporting the number of cells.
 
             </div>
 
             </div>
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             <div class="split_small"></div>
 
             <div class="split_small"></div>
 
             <div class="fb_48">
 
             <div class="fb_48">
−
                 <b>BBa_K3137005</b>
+
                 <b>BBa_K3137005 : <a href="http://parts.igem.org/Part:BBa_K3137005" style="color:blue;"alt=""><i>gntR</i></a></b>
 
             </div>
 
             </div>
 
             <br/>
 
             <br/>
 
             <div class="fm_22">
 
             <div class="fm_22">
−
                 <i>gntR</i>      996bp<br/>
+
                 996bp<br/>
−
                 The Gluconate repressor <i>gntR</i>, is a transcription factor that negatively regulates the operon
+
                 One of the most abundant and widely distributed groups of Helix-turn-helix (HTH) transcription factors is the metabolite-responsive GntR family of regulators. These proteins contain a DNA-binding HTH domain at the N-terminus of the protein and an effector-binding and/or oligomerisation domain at the C-terminus, where upon on binding an effector molecule, a conformational change occurs in the protein which influences the DNA-binding properties of the regulator resulting in repression or activation of transcription[1].  
−
                involved in
+
−
                the catabolism of d-gluconate via the <i>Entner-Doudoroff</i> pathway and represses genes involved in
+
−
                the
+
−
                different systems related to d-gluconate uptake: gluconate I and gluconate II.
+
 
             </div>
 
             </div>
  
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             <div class="split_small"></div>
 
             <div class="split_small"></div>
 
             <div class="fb_48">
 
             <div class="fb_48">
−
                 <b>BBa_K3137006</b>
+
                 <b>BBa_K3137006 : <a href="http://parts.igem.org/Part:BBa_K3137006" style="color:blue;"alt=""><i>abpAB</i></a></b>
 
             </div>
 
             </div>
 
             <br/>
 
             <br/>
 
             <div class="fm_22">
 
             <div class="fm_22">
−
                 <i>abpAB</i>      3803bp resistant protein<br/>
+
                 3803bp anti-protein<br/>
−
                 <i>AbpAB</i> are two genes of the <i>E.coli</i> genome that express resistant proteins that are
+
                 Gene <i>abpAB</i> is from <i>E.coli</i> genome which express anti-proteins that are resistant to T2,T4,T7
−
                resistant to T2,T4,T7
+
                 and λ phages. We obtained <i>abpA</i> and <i>abpB</i> by PCR from the genome of <i>E.coli</i> BL21 and
−
                 and λ phages. We obtained <i>abpA</i> and <i>abpB</i> by PCR from the genome of ?<i>E.coli</i> BL21 and
+
 
                 constructed
 
                 constructed
−
                 recombinant plasmid that connected <i>abpA</i> and <i>abpB</i> at the same time, then used IPTG to
+
                 recombinant plasmid that connected <i>abpA</i> and <i>abpB</i> at the same time. Then we used IPTG to
 
                 induce the
 
                 induce the
−
                 expression of resistant protein and verified the resistant function of the protein.
+
                 expression of anti-protein and verified the resistant function of the protein.[2]
 
             </div>
 
             </div>
  
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             <div class="split_small"></div>
 
             <div class="split_small"></div>
 
             <div class="fb_48">
 
             <div class="fb_48">
−
                 <b>BBa_K3137007</b>
+
                 <b>BBa_K3137007 : <a href="http://parts.igem.org/Part:BBa_K3137007" style="color:blue;"alt=""><i>rzpD</i> </a></b>
 
             </div>
 
             </div>
 
             <br/>
 
             <br/>
 
             <div class="fm_22">
 
             <div class="fm_22">
−
                 <i>rzpD</i>      462bp<br/>
+
                 462bp<br/>
−
                 Overexpression of <i>rzpD</i> causes abnormal biofilm architecture.
+
                 Overexpression of <i>rzpD</i> causes an abnormal biofilm architecture.
 
             </div>
 
             </div>
  
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             <div class="split_small"></div>
 
             <div class="split_small"></div>
 
             <div class="fb_48">
 
             <div class="fb_48">
−
                 <b>BBa_K3137008</b>
+
                 <b>BBa_K3137008 : <a href="http://parts.igem.org/Part:BBa_K3137008" style="color:blue;"alt=""><i>yhjH</i> </a></b>
 
             </div>
 
             </div>
 
             <br/>
 
             <br/>
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             <div class="split_small"></div>
 
             <div class="split_small"></div>
 
             <div class="fb_48">
 
             <div class="fb_48">
−
                 <b>BBa_K3137009</b>
+
                 <b>BBa_K3137009 : <a href="http://parts.igem.org/Part:BBa_K3137009" style="color:blue;"alt=""><i>nuoE</i></a></b>
 
             </div>
 
             </div>
 
             <br/>
 
             <br/>
 
             <div class="fm_22">
 
             <div class="fm_22">
−
                 <i>nuoE</i>      501bp<br/>
+
                 501bp<br/>
 
                 <i>NuoE</i> is part of the soluble fragment of NADH dehydrogenase I, which represents the electron input
 
                 <i>NuoE</i> is part of the soluble fragment of NADH dehydrogenase I, which represents the electron input
 
                 part
 
                 part
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             <div class="split_small"></div>
 
             <div class="split_small"></div>
 
             <div class="fb_48">
 
             <div class="fb_48">
−
                 <b>BBa_K3137010</b>
+
                 <b>BBa_K3137010 : <a href="http://parts.igem.org/Part:BBa_K3137010" style="color:blue;"alt="">T7 terminator</a></b>
 
             </div>
 
             </div>
 
             <br/>
 
             <br/>
 
             <div class="fm_22">
 
             <div class="fm_22">
−
                 T7 terminator      48bp terminator<br/>
+
                 48bp terminator<br/>
 
                 T7 terminator is used to quantify the level of expression in <i>E. coli</i>.
 
                 T7 terminator is used to quantify the level of expression in <i>E. coli</i>.
 
             </div>
 
             </div>
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             <div class="split_small"></div>
 
             <div class="split_small"></div>
 
             <div class="fb_48">
 
             <div class="fb_48">
−
                 <b>BBa_K3137011</b>
+
                 <b>BBa_K3137011 : <a href="http://parts.igem.org/Part:BBa_K3137011" style="color:blue;"alt=""><i>gfp</i></a></b>
 
             </div>
 
             </div>
 
             <br/>
 
             <br/>
 
             <div class="fm_22">
 
             <div class="fm_22">
−
                 <i>gfp</i>      717bp reporter<br/>
+
                 717bp reporter gene<br/>
−
                 The <i>gfp</i> can express green fluorescent protein, which can be used as a marker gene in the process
+
                 The <i>gfp</i> can express green fluorescent protein, which can be used as a reporter gene in the process
−
                 to report the number of cells.
+
                 of reporting the number of cells.
 
             </div>
 
             </div>
  
−
            <!--            BBa_K628000-->
+
         
−
            <div class="split_small"></div>
+
−
            <div class="fb_48">
+
−
                <b>BBa_K3137011</b>
+
−
            </div>
+
−
            <br/>
+
−
            <div class="fm_22">
+
−
                codes for<i>protegrin-1</i>      (anti-microbial peptide) 60bp
+
−
                antibacterial peptides<br/>
+
−
                <i>Protegrin-1</i> coding region. <i>Protegrin-1</i> is an '18-residue beta-sheet peptide isolated from
+
−
                porcine
+
−
                leukocytes with antimicrobial activity against a broad range of microorganisms.' It has its effect by
+
−
                pore membrane disruption and possibly also by effects such as activation of membrane-damaging proteases
+
−
                and has anti-microbial activity against <i>E. coli</i>.
+
−
            </div>
+
  
  
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             <div class="split_small"></div>
 
             <div class="split_small"></div>
 
             <div class="fb_48">
 
             <div class="fb_48">
−
                 <b>BBa_K3137013</b>
+
                 <b>BBa_K3137013 : <a href="http://parts.igem.org/Part:BBa_K3137013" style="color:blue;"alt="">report circuit</a></b>
 
             </div>
 
             </div>
 
             <br/>
 
             <br/>
 
             <div class="fm_22">
 
             <div class="fm_22">
−
                report circuit        1666bp
+
            1666bp
 
             </div>
 
             </div>
  
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                 style="width: 100%;height: auto;" alt="1666bp">
 
                 style="width: 100%;height: auto;" alt="1666bp">
 
             <div class="fm_22">
 
             <div class="fm_22">
−
                 The report gene circuit consists of two periods of phage infection. In incubation period (about 5 min),
+
                  
−
                there is an inducible promoter <i>PputA</i> and a green fluorescent protein gene <i>gfp</i>. In outbreak
+
The report gene circuit consists of two periods of phage infection. In incubation period (about 5 min), there is an inducible promoter PputA and a green fluorescent protein gene <i>gfp</i>. In burst period (about 20 min), there is an inducible promoter PglcF, and a red fluorescent protein gene <i>mCherry</i>.
−
                period (about
+
 
−
                20 min), there is an inducible promoter <i>PglcF</i>, and a red fluorescent protein gene <i>mCherry</i>.
+
When <i>E. coli</i> is infected by a phage, report gene circuit is induced to express. When a phage infects <i>E. coli</i> into the incubation period (about 5 min), the inducible promoter PputA will induce the bacteria to expressthe green fluorescent protein gene <i>gfp</i>.
−
                When <i>E. coli</i> is infected by a phage, both report and respond gene circuits are induced to express
+
 
−
                at the
+
If the resistant proteins successfully resist phage infection, the inducible promoter PglcF will not induce the expression of downstream gene. If the resistant proteins fail to kill phages, it means that phages will continue to infect bacteria, when it comes to the burst period (around 20 min) of phage infection, the inducible promoter PglcF will activate the red fluorescent protein gene <i>mCherry</i>. so that the bacteria will display red fluoresce and lyse cells before the assembly of phages.
−
                same time. When a phage infects <i>E. coli</i> into the incubation period (about 5 min), the inducible
+
 
−
                promoter
+
 
−
                <i>PputA</i> will induce the bacteria to express the resistant protein and the green fluorescent protein
+
−
                gene
+
−
                <i>gfp</i>.
+
−
                If the resistant protein successfully resists phage infection, the inducible promoter <i>PglcF</i> will
+
−
                not
+
−
                induce the expression of downstream gene. If the resistant protein fails to kill phages, it means that
+
−
                phages will continue to infect bacteria, when it comes to the outbreak period (around 20 min) of phage
+
−
                infection, the inducible promoter <i>PglcF</i> will activate the red fluorescent protein gene
+
−
                <i>mCherry</i> and the
+
−
                expression of downstream toxic protein gene so that the bacteria will display red fluoresce and lyse
+
−
                cells before the assembly of phages.
+
 
             </div>
 
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                 <b>BBa_K3137014</b>
+
                 <b>BBa_K3137014 : <a href="http://parts.igem.org/Part:BBa_K3137014" style="color:blue;"alt="">response circuit</a></b>
 
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             <br/>
 
             <div class="fm_22">
 
             <div class="fm_22">
−
                respond circuit        5097bp
+
            5097bp
 
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                 The respond gene circuit is composed of an inducible promoter <i>PputA</i> and anti-phage protein genes of the
+
                 The responce gene circuit is composed of an inducible promoter PputA and anti-phage protein genes of the incubation period (about 5 min), and an inducible promoter PglcF and a toxic protein gene protegrin-1 of the burst period (about 20 min).
−
                incubation period (about 5 min), and an inducible promoter <i>PglcF</i> and a toxic protein gene <i>protegrin-1</i> of
+
 
−
                the outbreak period (about 20 min).
+
When <i>E. coli</i> is infected by a phage, responce gene circuit is induced to express. When a phage infects <i>E. coli</i> into the incubation period (about 5 min), the inducible promoter PputA will induce the bacteria to express the resistant proteins.
−
                When <i>E. coli</i> is infected by a phage, both report and respond gene circuits are induced to express at the
+
 
−
                same time. When a phage infects <i>E. coli</i> into the incubation period (about 5 min), the inducible promoter
+
If the resistant proteins successfully resist phage infection, the inducible promoter PglcF will not induce the expression of downstream gene. If the resistant proteins fail to kill phages, it means that phages will continue to infect bacteria, when it comes to the burst period (around 20 min) of phage infection, the inducible promoter PglcF will activate the expression of downstream toxic protein gene so that the bacteria will lyse cells before the assembly of phages.
−
                <i>PputA</i> will induce the bacteria to express the resistant protein <i>abpAB</i> and <i>gntR</i>, and the green
+
 
−
                fluorescent protein gene <i>gfp</i>.
+
−
                If the resistant proteins successfully resist phage infection, the inducible promoter <i>PglcF</i> will not
+
−
                induce the expression of downstream gene. If the resistant proteins fail to kill phages, it means that
+
−
                phages will continue to infect bacteria, when it comes to the outbreak period (around 20 min) of phage
+
−
                infection, the inducible promoter <i>PglcF</i> will activate the red fluorescent protein gene <i>mCherry</i> and the
+
−
                expression of downstream toxic protein gene <i>protegrin-1</i> so that the bacteria will display red fluoresce
+
−
                and lyse cells before the assembly of phages.
+
 
             </div>
 
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             <img src="https://static.igem.org/mediawiki/2019/2/2f/T--JiangnanU_China--parts_4.png"
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             <img src="https://static.igem.org/mediawiki/2019/b/bb/T--JiangnanU_China--parts--12138.png"
 
                 style="width: 100%;height: auto;">
 
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             <img src="https://static.igem.org/mediawiki/2019/d/d9/T--JiangnanU_China--parts_3.png"
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             <img src="https://static.igem.org/mediawiki/2019/8/81/T--JiangnanU_China--parts_2000.png"
 
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 +
 +
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<div class="fm_22">
 +
 +
                References[1-2]
 +
                <br/>
 +
                <br/>
 +
                [1] Hoskisson P A , Sébastien Rigali. Chapter 1: Variation in form and function the helix-turn-helix regulators of the GntR superfamily[J]. Advances in applied microbiology, 2009, 69(69):1-22.
 +
                <br />         
 +
[2] Yasui R, Washizaki A, Furihata Y, Yonesaki T, Otsuka Y: AbpA and AbpB provide anti-phage activity in <i>Escherichia coli</i>. Genes Genet Syst 2014, 89(2):51-60.
 +
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Latest revision as of 02:26, 22 October 2019

JiangNan

Basic Parts
BBa_K3137000 : PrsmH

250bp constitutive promoter
We ligated promoter rsmH with gfp to measure the fluorescence intensity of GFP.
BBa_K3137001 : Improved RBS 4

11bp RBS
As we all know, RBS refers to a sputum-rich untranslated region upstream of the initiation codon AUG. BBa_B0033 is a weaker RBS based on Ron Weiss thesis. Compared with BBa_B0030, BBa_B0031, BBa_B0032, BBa_B0034, BBa_B0033 is the weakest, and whose RBS strength is 0.35% of BBa_B0034. By analyzing their sequences, we found that BBa_B0033 has fewer purines and no not have AAA sequence. SO we designed the RBS by adjusting the proportion of bases.
Then we ligated the two RBS with promotor rsmH (BBa_K3137000) and gene gfp (BBa_K3137011) and transform into E. coli BL21 to compare their green fluorescence. And take E. coli BL21 as a control. The figure below shows that BBa_K3137001 has the stronger fluorescently protein expression than BBa_B0033, that means it’s efficient to increase the protein expression by increasing the number of purines in the RBS sequence, which is beneficial to the binding of ribosomes.
BBa_K3137002 : PputA

250bp inducible promoter
PputA is an inducible promoter. When E.coli is infected by a phage, both report and response gene circuits are induced to express at the same time. When a phage infects Escherichia coli into the latent period (about 5 min), PputA will induce the bacteria to express anti-protein and display green fluorescence.
BBa_K3137003 : PglcF

250bp inducible promoter
PglcF is an inducible promoter that works when phages infect bacterial for around 20 min. If the anti-proteins successfully resist phage infection, the inducible promoter PglcF will not induce the expression of downstream gene. If the anti-proteins cannot kill phages, the phages will continue to infect. When phages infect bacteria into the burst period (about 20 min), the inducible promoter PglcF will induce expression of the red fluorescent protein gene mCherry and the downstream toxic protein gene protegrin-1, which will make the bacteria display red fluorescence and lyse cells before the assembly of phages.
BBa_K3137004 : mCherry

708bp reporter gene
The mCherry can express red fluorescent protein and it is used as a reporter gene in the process of reporting the number of cells.
BBa_K3137005 : gntR

996bp
One of the most abundant and widely distributed groups of Helix-turn-helix (HTH) transcription factors is the metabolite-responsive GntR family of regulators. These proteins contain a DNA-binding HTH domain at the N-terminus of the protein and an effector-binding and/or oligomerisation domain at the C-terminus, where upon on binding an effector molecule, a conformational change occurs in the protein which influences the DNA-binding properties of the regulator resulting in repression or activation of transcription[1].
BBa_K3137006 : abpAB

3803bp anti-protein
Gene abpAB is from E.coli genome which express anti-proteins that are resistant to T2,T4,T7 and λ phages. We obtained abpA and abpB by PCR from the genome of E.coli BL21 and constructed recombinant plasmid that connected abpA and abpB at the same time. Then we used IPTG to induce the expression of anti-protein and verified the resistant function of the protein.[2]
BBa_K3137007 : rzpD

462bp
Overexpression of rzpD causes an abnormal biofilm architecture.
BBa_K3137008 : yhjH

yhjH 768bp
YhjH protein contains a EAL domain to catalyze c-di-GMP into GMP, so as to regulate the levels of c-di-GMP.
BBa_K3137009 : nuoE

501bp
NuoE is part of the soluble fragment of NADH dehydrogenase I, which represents the electron input part of the enzyme.
BBa_K3137010 : T7 terminator

48bp terminator
T7 terminator is used to quantify the level of expression in E. coli.
BBa_K3137011 : gfp

717bp reporter gene
The gfp can express green fluorescent protein, which can be used as a reporter gene in the process of reporting the number of cells.
Composite Part
BBa_K3137013 : report circuit

1666bp
1666bp
The report gene circuit consists of two periods of phage infection. In incubation period (about 5 min), there is an inducible promoter PputA and a green fluorescent protein gene gfp. In burst period (about 20 min), there is an inducible promoter PglcF, and a red fluorescent protein gene mCherry. When E. coli is infected by a phage, report gene circuit is induced to express. When a phage infects E. coli into the incubation period (about 5 min), the inducible promoter PputA will induce the bacteria to expressthe green fluorescent protein gene gfp. If the resistant proteins successfully resist phage infection, the inducible promoter PglcF will not induce the expression of downstream gene. If the resistant proteins fail to kill phages, it means that phages will continue to infect bacteria, when it comes to the burst period (around 20 min) of phage infection, the inducible promoter PglcF will activate the red fluorescent protein gene mCherry. so that the bacteria will display red fluoresce and lyse cells before the assembly of phages.
BBa_K3137014 : response circuit

5097bp
5097bp
The responce gene circuit is composed of an inducible promoter PputA and anti-phage protein genes of the incubation period (about 5 min), and an inducible promoter PglcF and a toxic protein gene protegrin-1 of the burst period (about 20 min). When E. coli is infected by a phage, responce gene circuit is induced to express. When a phage infects E. coli into the incubation period (about 5 min), the inducible promoter PputA will induce the bacteria to express the resistant proteins. If the resistant proteins successfully resist phage infection, the inducible promoter PglcF will not induce the expression of downstream gene. If the resistant proteins fail to kill phages, it means that phages will continue to infect bacteria, when it comes to the burst period (around 20 min) of phage infection, the inducible promoter PglcF will activate the expression of downstream toxic protein gene so that the bacteria will lyse cells before the assembly of phages.
References[1-2]

[1] Hoskisson P A , Sébastien Rigali. Chapter 1: Variation in form and function the helix-turn-helix regulators of the GntR superfamily[J]. Advances in applied microbiology, 2009, 69(69):1-22.
[2] Yasui R, Washizaki A, Furihata Y, Yonesaki T, Otsuka Y: AbpA and AbpB provide anti-phage activity in Escherichia coli. Genes Genet Syst 2014, 89(2):51-60.
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