Difference between revisions of "Part:BBa K1332010"
 
(7 intermediate revisions by 2 users not shown)
Line 5: Line 5:
  
 
<h3>explanation of this part</h3>
 
<h3>explanation of this part</h3>
−
[[File:Gifu 5' not remove2.png|500px|]]<br>
+
[[File:Gifu expla doubt.png|500px|]]<br>
−
<b>Figure 1. mRNA circularization device (3'side)(not endless translation)</b><br><br>
+
<b>Figure 1. mRNA circularization device (3'side)(not endless translation)</b><br>
 +
<b>Link:</b>
 +
<ul>
 +
<li>[https://parts.igem.org/Part:BBa_K1332004 BBa_K1332004]:The 5' side of the intron(+exon fragment) from td gene of T4 phage</li>
 +
<li>[https://parts.igem.org/Part:BBa_K1332005 BBa_K1332005]:The 3' side of the intron(+exon fragment) from td gene of T4 phage</li>
 +
<li>[https://parts.igem.org/Part:BBa_K1332008 BBa_K1332008]:mRNA circularization device (5´ side)</li>
 +
<li>[https://parts.igem.org/Part:BBa_K1332010 BBa_K1332010]:mRNA circularization device (3´ side)</li>
 +
</ul>
 +
<br><br>
 
Part which surrounded by red closing line is <b>mRNA circularization device (3’side)(not endless translation)</b>.(figure 1)<br>
 
Part which surrounded by red closing line is <b>mRNA circularization device (3’side)(not endless translation)</b>.(figure 1)<br>
 
This part has a stop codon, so a circular mRNA made by this part <b>doesn’t cause continuous translation</b>.  
 
This part has a stop codon, so a circular mRNA made by this part <b>doesn’t cause continuous translation</b>.  
Line 17: Line 25:
  
 
<h1><b>When protein coding is RFP coding, its results shows as follow.</b></h1>
 
<h1><b>When protein coding is RFP coding, its results shows as follow.</b></h1>
−
[[File:Gifu RFP.png|500px|]]<br>
+
[[File:Gifu 10 RFP occasion.png|500px|]]<br>
 
<b>Figure 2.In the case that RFP is inserted as protein coding. </b><br>
 
<b>Figure 2.In the case that RFP is inserted as protein coding. </b><br>
−
An RFP which we use is [https://parts.igem.org/Part:BBa_K1332002 BBa_K1332002].
+
<b>Link:</b>
−
That's an RFP which combines with a Histidine tag.
+
<ul>
−
 
+
<li>An RFP which we use is [https://parts.igem.org/Part:BBa_K1332002 BBa_K1332002].
−
 
+
That's an RFP which combines with a Histidine tag.</li>
−
 
+
</ul>
−
 
+
  
  
Line 36: Line 43:
 
[[File:gifu sds remove 3.png|500px|]]<br>
 
[[File:gifu sds remove 3.png|500px|]]<br>
  
−
<b>Figure 3. The result of SDS-PAGE</b>
+
<b>Figure 3. The result of SDS-PAGE</b><br>
 +
In the case that this part was used {Circular + RFP with stop codon (5’ intron)}, long-chain protein was not synthesized.
 +
 
  
  

Latest revision as of 20:41, 2 November 2014

mRNA circularization device (3 side)

This part consists of the 5' side of the intron(+exon fragment) from td gene of T4 phage(BBa_K1332004) and a double terminator(BBa_B0015). The protein coding sequence that is inserted between this device and mRNA circularization device (5’ side)(BBa_K1332008) can be circularized. But the 5' side of the intron(+exon fragment) from td gene of T4 phage contains a stop codon, so a circular mRNA made by this part doesn’t cause continuous translation.

explanation of this part

Gifu expla doubt.png
Figure 1. mRNA circularization device (3'side)(not endless translation)
Link:

  • BBa_K1332004:The 5' side of the intron(+exon fragment) from td gene of T4 phage
  • BBa_K1332005:The 3' side of the intron(+exon fragment) from td gene of T4 phage
  • BBa_K1332008:mRNA circularization device (5´ side)
  • BBa_K1332010:mRNA circularization device (3´ side)



Part which surrounded by red closing line is mRNA circularization device (3’side)(not endless translation).(figure 1)
This part has a stop codon, so a circular mRNA made by this part doesn’t cause continuous translation.

How to use

Insert the protein coding sequence between this device and mRNA circularization device (5’ side). How to use this part is same as how to use 5' side device(BBa_K1332008) or 3' side device(endless translation)(BBa_K1332009) . Thus,read that.


When protein coding is RFP coding, its results shows as follow.

Gifu 10 RFP occasion.png
Figure 2.In the case that RFP is inserted as protein coding.
Link:

  • An RFP which we use is BBa_K1332002. That's an RFP which combines with a Histidine tag.


Synthesis of long-chain proteins

Summary of the experiment

Confirm repeating translation by SDS-PAGE([http://2014.igem.org/Team:Gifu/Protocol#SDS protocol]).


Result

Gifu sds remove 2.png
Gifu sds remove 3.png

Figure 3. The result of SDS-PAGE
In the case that this part was used {Circular + RFP with stop codon (5’ intron)}, long-chain protein was not synthesized.



Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]