Difference between revisions of "Part:BBa K3050000"
Line 5: Line 5:
 
Magainin-2 expressed inside E.coli combines to cell membrane and damages it.In our project, we aimed to make BL21(DE3) lyse easier by transforming BBa_K3050000 on pSB1C3.
 
Magainin-2 expressed inside E.coli combines to cell membrane and damages it.In our project, we aimed to make BL21(DE3) lyse easier by transforming BBa_K3050000 on pSB1C3.
  
−
<!-- Add more about the biology of this part here
 
 
===Usage and Biology===
 
===Usage and Biology===
  
Line 18: Line 17:
 
GFP part is on downstream of T7 promotor and magainin-2, hence we're quite sure that this result indicates that magainin-2 is expressed at the same time.  
 
GFP part is on downstream of T7 promotor and magainin-2, hence we're quite sure that this result indicates that magainin-2 is expressed at the same time.  
  
−
<div class="students">
+
[[File:T--Gunma--BBa_K3050000.png|600px|center|]]
−
    <div class="card">
+
Fig.1 BL21(DE3) pSB1C3 BBa_K3050000 expressed under visible light with negative control without IPTG induction
−
        <img src="https://2019.igem.org/wiki/images/b/b9/T--Gunma--BBa_K3050000.png">
+
−
    </div>
+
−
</div>Fig.1 BL21(DE3) pSB1C3 BBa_K3050000 expressed under visible light with negative control without IPTG induction
+
  
  
 +
[[File:T--Gunma--BBa_K3050000_2.png|600px|center|]]
 +
Fig.2 pSB1C3 BBa_K3050000 expressed under UV light with negative control without IPTG induction
  
−
<div class="clear"></div>
 
  
−
<div class="students">
+
[[File:T--Gunma--BBa_K3050000_3.png|600px|center|]]
−
    <div class="card">
+
Fig.3 Four biological replicates all highly expressed
−
        <img src="https://2019.igem.org/wiki/images/e/e2/T--Gunma--BBa_K3050000_2.png">
+
−
    </div>
+
−
</div>Fig.2 pSB1C3 BBa_K3050000 expressed under UV light with negative control without IPTG induction
+
  
  
−
<div class="clear"></div>
 
  
−
<div class="students">
 
−
    <div class="card">
 
−
        <img src="https://2019.igem.org/wiki/images/0/05/T--Gunma--BBa_K3050000_3.png">
 
−
    </div>
 
−
</div>Fig.3 Four biological replicates all highly expressed
 
−
 
−
<div class="clear"></div>
 
−
 
−
<!-- -->
 
 
<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
 
<partinfo>BBa_K3050000 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3050000 SequenceAndFeatures</partinfo>

Revision as of 19:29, 21 October 2019


pT7+RBS+mag-2+GFP+double_terminator

Magainin-2 expressed inside E.coli combines to cell membrane and damages it.In our project, we aimed to make BL21(DE3) lyse easier by transforming BBa_K3050000 on pSB1C3.

Usage and Biology

This is a composite part that includes T7 promotor + RBS + mag-2 + GFP + double terminators.

BBa_I746909 is linerized by Inverse PCR to be combined with magainin-2 sequense by In-Fusion reaction. https://parts.igem.org/Part:BBa_I746909

BL21(DE3) transformed pSB1C3 BBa_K3050000 successfully grew and emit strong green fluorescent light as shown in pictures. Thus we succeeded to prove Inverse PCR,In-Fusion reaction,and transformation went great.

GFP part is on downstream of T7 promotor and magainin-2, hence we're quite sure that this result indicates that magainin-2 is expressed at the same time.

T--Gunma--BBa K3050000.png

Fig.1 BL21(DE3) pSB1C3 BBa_K3050000 expressed under visible light with negative control without IPTG induction


T--Gunma--BBa K3050000 2.png

Fig.2 pSB1C3 BBa_K3050000 expressed under UV light with negative control without IPTG induction


T--Gunma--BBa K3050000 3.png

Fig.3 Four biological replicates all highly expressed


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
    INCOMPATIBLE WITH RFC[1000]
    Illegal SapI.rc site found at 190