Difference between revisions of "Part:BBa K3050000"

 
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<partinfo>BBa_K3050000 short</partinfo>
 
<partinfo>BBa_K3050000 short</partinfo>
  
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Magainin-2 expressed inside E.coli combines to cell membrane and damages it.In our project, we used it to weaken the cell membrane to make it easier to lyse after cell wall is broken by T7 lysozyme in BL21(DE3)pLysS.
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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.
  
 
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===Usage and Biology===
 
===Usage and Biology===
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This is a composite part that includes T7 promotor + RBS + mag-2 + GFP + double terminators.
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BBa_I746909 is linerized by Inverse PCR to be combined with magainin-2 sequense by In-Fusion reaction.
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https://parts.igem.org/Part:BBa_I746909
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BL21(DE3) transformed pSB1C3 BBa_K3050000 successfully grew and emit strong green fluorescent light as shown in pictures.
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Thus we succeeded to prove Inverse PCR,In-Fusion reaction,and transformation went great.
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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.
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<div class="students">
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    <div class="card">
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        <img src="https://2019.igem.org/wiki/images/b/b9/T--Gunma--BBa_K3050000.png">
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    </div>
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</div>Fig.1 BL21(DE3) pSB1C3 BBa_K3050000 expressed under visible light with negative control without IPTG induction
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<div class="clear"></div>
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<div class="students">
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    <div class="card">
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        <img src="https://2019.igem.org/wiki/images/e/e2/T--Gunma--BBa_K3050000_2.png">
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    </div>
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</div>Fig.2 pSB1C3 BBa_K3050000 expressed under UV light with negative control without IPTG induction
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<div class="clear"></div>
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<div class="students">
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    <div class="card">
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        <img src="https://2019.igem.org/wiki/images/0/05/T--Gunma--BBa_K3050000_3.png">
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    </div>
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</div>Fig.3 Four biological replicates all highly expressed
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<div class="clear"></div>
  
 
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Revision as of 19:19, 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.

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