View source for Part:BBa K5323995


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<partinfo>BBa_K5323995 short</partinfo>

Uses a factor strong RBS(BBa_J34801), AFE_1379 (BBa_K5323998). This part is an easy BioBrick.

===Usage and Biology===
This component is used to overexpress DGC in the engineering bacterium <i>Acidithiobacillus ferrooxidans</i> and participate in screening the most efficient cyclic diguanylate dynamic synthesis strains.
We cloned the highly efficient DGC yedQ in <i>E.coli</i> and DGC AFE_0053, AFE_1373, AFE_1379 in <i>A. ferrooxidans</i> genome into the expression vector pYDT to construct the highly efficient expression vector of DGC. Furthermore, the carrier is conjured into <i>A. ferrooxidans</i>, the concentration of biofilm and c-di-GMP is measured, and the most efficient dynamic regulatory gene of c-di-GMP is screened.
<center>https://static.igem.wiki/teams/5323/parts/fig7.png</center>
<center>Fig 1 <i>Acidithiobacillus ferrooxidans</i> pore plate biofilm formation column</center>
<center>https://static.igem.wiki/teams/5323/parts/fig8.png</center>
<center>Fig 2  <i>Escherichia coli</i>  pore plate biofilm formation column</center>
<center>https://static.igem.wiki/teams/5323/parts/fig9.png</center>
<center>Fig 3 Schematic diagram of c-di-GMP content of <i>E.coli</i> BL21 YDT, AFE_1373, AFE_1379, YedQ. A is the c-di-GMP content of <i>E.coli</i> BL21 YDT, AFE_1373, AFE_1379, YedQ. </center>
<center>https://static.igem.wiki/teams/5323/parts/fig10.png</center>
<center>Fig 4 indicates the standard curve of c-di-GMP</center>
Since it is previously known that IPTG can promote the significant expression of biofilm, after obtaining the growth characteristics of <i>A. ferrooxidans</i>, we directly measure the biofilm content in different time periods when the inducer IPTG is added to the strain. AFE_1379 was conducive to biofilm formation, and its significant difference P < 0.05 also indicated that its significantly promoted biofilm formation (Fig 3). In addition, we also determined the c-di-GMP content of <i>E.coli</i> BL21 transferred into plasmids pAFE_1373, pAFE_1379, pYedQ, and no-load pYDT. We calculated the c-di-GMP content according to the standard curve (Fig 4). With the addition of inducer IPTG, the C-di-GMP contents of pAFE_1373, pAFE_1379, pYedQ were significantly increased except for no-load plasmids.
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<span class='h3bb'>Sequence and Features</span>
<partinfo>BBa_K5323995 SequenceAndFeatures</partinfo>


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===Functional Parameters===
<partinfo>BBa_K5323995 parameters</partinfo>
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