Difference between revisions of "Part:BBa K4325022"
 
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__NOTOC__
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<partinfo>BBa_K4325022 short</partinfo>
 
<partinfo>BBa_K4325022 short</partinfo>
 
===Description===
 
===Description===
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The composite part is a generator consisting of pDawn (cI-LVA) (<partinfo>BBa_K1075044</partinfo>) and X174E-LVA (<partinfo>BBa_K4325007</partinfo>).
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The composite part is a generator consisting of pDawn(cI-LVA) (<partinfo>BBa_K1075044</partinfo>), X174E-LVA (<partinfo>BBa_K4325007</partinfo>) and T1 terminator (<partinfo>BBa_K3033016</partinfo>).
 
===Usage===
 
===Usage===
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<p>We inserted the pDawn (cI-LVA)(<partinfo>BBa_K1075044</partinfo>) blue light response system and the lysis gene X174E-LVA(<partinfo>BBa_K4325007</partinfo>) into the pSEVA331 expression vector, which was inserted into <i>E.coli</i> TOP10 and screened out the bacterial colony which grew in the dark but did not grow under the blue light to verify the responsiveness of pDawn (cI-LVA) to blue light.</p>
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<p>We inserted the blue light response system pDawn(cI-LVA) (<partinfo>BBa_K1075044</partinfo>) and the lysis gene X174E-LVA (<partinfo>BBa_K4325007</partinfo>) into the pSEVA331 expression vector, which was introduced into <i>E.coli</i> TOP10 and screened for the bacterial colonies which grew in the dark but did not grow under blue light to verify the responsiveness of pDawn(cI-LVA) to blue light.</p>
  
 
===Sequence and Features===
 
===Sequence and Features===
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=2022 SZPT-China=
 
=2022 SZPT-China=
 
<h3>Characterization</h3>
 
<h3>Characterization</h3>
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<h4>1.Batch screening of pDawn(cI-LVA)-X174E-LVA-T1 in response to blue light lysis in <i>E. coli</i>.</h4>
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<h4>1.Batch screening of pDawn-X174E-LVA-T1 by evaluating the response to blue light.</h4>
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<p>We transformed pDawn(cI-LVA)-X174E-LVA-T1 into TOP10 and performed agarose gel electrophoresis, as shown in Figure 1,pDawn(cI-LVA)-X174E-LVA-T1 has been successfully transformed into TOP10.</p>
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<p>As shown in Figure 1, we successfully introduced pSEVA331-pDawn(cI-LVA)-X174E-LVA-T1 into <i>E.coli</i> TOP10.</p>
  
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[[File:K18 1.png|600px|thumb|center|Figure 1: <p></p>pDawn (cI-LVA)-RBS070-LKD-LVA-T1;and (2)pSEVA331-pDawn(cI-LVA)-X174E-LVA-T1 has been successfully identified in TOP10]]
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[[File:K18 1.png|600px|thumb|center|Figure 1: (1) pDawn(cI)-X174E-T1; <b>(2) pDawn(cI-LVA)-X174E-LVA-T1</b>; (3) pDawn(cI)-LKD-T1; (4) pDawn(cI-LVA)-RBS070-LKD-LVA-T1 were constructed successfully.]]
  
 
<h3>References</h3>
 
<h3>References</h3>
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<p>[1]Robert Ohlendorf, Roee R. Vidavski, Avigdor Eldar et.al.From Dusk till Dawn: One-Plasmid Systems for Light-Regulated Gene Expression.Journal of Molecular Biology, 08 Jan 2012, 416(4):534-542.</p>
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<p>[1] Ohlendorf, R., Vidavski, R. R., Eldar, A., Moffat, K. & Möglich, A. From Dusk till Dawn: One-Plasmid Systems for Light-Regulated Gene Expression. J. Mol. Biol. 416, 534-542 (2012).</p>
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<p>[2] Witte, A., Wanner, G., Sulzner, M. & Lubitz, W. Dynamics of PhiX174 protein E-mediated lysis of <i>Escherichia coli</i>. Arch. Microbiol. 157, 381-8 (1992).</p>

Latest revision as of 14:37, 12 October 2022

pDawn-X174E-LVA-T1

Description

The composite part is a generator consisting of pDawn(cI-LVA) (BBa_K1075044), X174E-LVA (BBa_K4325007) and T1 terminator (BBa_K3033016).

Usage

We inserted the blue light response system pDawn(cI-LVA) (BBa_K1075044) and the lysis gene X174E-LVA (BBa_K4325007) into the pSEVA331 expression vector, which was introduced into E.coli TOP10 and screened for the bacterial colonies which grew in the dark but did not grow under blue light to verify the responsiveness of pDawn(cI-LVA) to blue light.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 2171
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 63
    Illegal NgoMIV site found at 195
    Illegal NgoMIV site found at 289
    Illegal NgoMIV site found at 582
    Illegal NgoMIV site found at 1076
    Illegal NgoMIV site found at 1094
    Illegal NgoMIV site found at 1184
    Illegal AgeI site found at 414
    Illegal AgeI site found at 1542
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1643
    Illegal BsaI.rc site found at 525


2022 SZPT-China

Characterization

1.Batch screening of pDawn-X174E-LVA-T1 by evaluating the response to blue light.

As shown in Figure 1, we successfully introduced pSEVA331-pDawn(cI-LVA)-X174E-LVA-T1 into E.coli TOP10.

Figure 1: (1) pDawn(cI)-X174E-T1; (2) pDawn(cI-LVA)-X174E-LVA-T1; (3) pDawn(cI)-LKD-T1; (4) pDawn(cI-LVA)-RBS070-LKD-LVA-T1 were constructed successfully.

References

[1] Ohlendorf, R., Vidavski, R. R., Eldar, A., Moffat, K. & Möglich, A. From Dusk till Dawn: One-Plasmid Systems for Light-Regulated Gene Expression. J. Mol. Biol. 416, 534-542 (2012).

[2] Witte, A., Wanner, G., Sulzner, M. & Lubitz, W. Dynamics of PhiX174 protein E-mediated lysis of Escherichia coli. Arch. Microbiol. 157, 381-8 (1992).