Difference between revisions of "Part:BBa K2308016"

 
Line 5: Line 5:
 
This part contains LacIq repressor, LacIq spacer, LacIq promotor and LacIq coding sequence and is used for Rhodobacter sphaeroides, cells with the device can carry out inducible expression of the target proteins.
 
This part contains LacIq repressor, LacIq spacer, LacIq promotor and LacIq coding sequence and is used for Rhodobacter sphaeroides, cells with the device can carry out inducible expression of the target proteins.
  
−
<!-- Add more about the biology of this part here
 
−
===Usage and Biology===
 
−
 
−
<!-- -->
 
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K2308016 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K2308016 SequenceAndFeatures</partinfo>
 +
<partinfo>BBa_K2308016 parameters</partinfo>
  
  
−
<!-- Uncomment this to enable Functional Parameter display
+
 
−
===Functional Parameters===
+
<html>
−
<partinfo>BBa_K2308016 parameters</partinfo>
+
−
<!-- -->
+
<body>
 +
 +
<p>sYFP2(BBa_K2308003) is the key part in our experiment. It is used both for fusion expression (see BBa_K2308002) and inducible cytoplasm expression (combined with BBa_K2308016), and the host cell is Rhodobacter sphaeroides 2.4.1. </p><br>
 +
<div class="row" align="left" style="margin-left:20%;">
 +
<img src="https://static.igem.org/mediawiki/2017/4/47/F1-D.png" style="width: 20%;">
 +
<img src="https://static.igem.org/mediawiki/2017/f/fa/F1-E.png" style="width: 20%;">
 +
</div>
 +
<center><p>Figure 1：PCR test of BBa_K2308003(optimized sYFP2) and original sYFP2</p></center><br><br>
 +
 
 +
 
 +
<p> In inducible cytoplasm expression experiment, we used part BBa_K2308016 to turn the <i>Rhodobacter sphaedoides 2.4.1</i>  into inducible strains, and IPTG was added （800 μM in final volume）when the OD700 of the strain was about 0.4（grown for about 24h）.</p>
 +
<div class="row" align="left" style="margin-left:20%;">
 +
<img src="https://static.igem.org/mediawiki/2017/thumb/c/c7/Kp6.png" style="width: 20%;">
 +
<img src="https://static.igem.org/mediawiki/2017/thumb/2/2e/Kp7.png" style="width: 20%;">
 +
</div>
 +
<center><p>Figure 2：fluorescent image of sYFP2 (original) and sYFP2(optimized)</p></center>
 +
<br><br>
 +
 
 +
<div class="row" align="left" style="margin-left:20%;">
 +
<img src="https://static.igem.org/mediawiki/2017/9/99/F5-A%281%29.png" style="width: 20%;">
 +
<img src="https://static.igem.org/mediawiki/2017/9/99/F5-A%281%29.png" style="width: 20%;">
 +
</div>
 +
<center><p>Figure 3：Fluorescence intensity of WT, sYFP2(optimized), sYFP2(unoptimized) strains.</p></center>
 +
<br><br>
 +
 
 +
<p> It is obvious that after the codon optimization, sYFP2 can be better expressed in Rhodobacter sphaeroides 2.4.1, and the LacIq promotor also worked very well.</p>
 +
 
 +
 
 +
 
 +
 
 +
</body>
 +
 
 +
</html>

Revision as of 10:33, 31 October 2017


LacIq

This part contains LacIq repressor, LacIq spacer, LacIq promotor and LacIq coding sequence and is used for Rhodobacter sphaeroides, cells with the device can carry out inducible expression of the target proteins.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal XhoI site found at 84
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]


sYFP2(BBa_K2308003) is the key part in our experiment. It is used both for fusion expression (see BBa_K2308002) and inducible cytoplasm expression (combined with BBa_K2308016), and the host cell is Rhodobacter sphaeroides 2.4.1.


Figure 1：PCR test of BBa_K2308003(optimized sYFP2) and original sYFP2



In inducible cytoplasm expression experiment, we used part BBa_K2308016 to turn the Rhodobacter sphaedoides 2.4.1 into inducible strains, and IPTG was added （800 μM in final volume）when the OD700 of the strain was about 0.4（grown for about 24h）.

Figure 2：fluorescent image of sYFP2 (original) and sYFP2(optimized)



Figure 3：Fluorescence intensity of WT, sYFP2(optimized), sYFP2(unoptimized) strains.



It is obvious that after the codon optimization, sYFP2 can be better expressed in Rhodobacter sphaeroides 2.4.1, and the LacIq promotor also worked very well.