Difference between revisions of "Part:BBa K4325020"
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[[File:K20 3.png|600px|thumb|center|Figure 3: Growth curve diagram of pSEVA331-pDawn(cI-LVA)-RBS070-X174E-T1-TOP10.]]
 
[[File:K20 3.png|600px|thumb|center|Figure 3: Growth curve diagram of pSEVA331-pDawn(cI-LVA)-RBS070-X174E-T1-TOP10.]]
  
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<h3>2.pSEVA331-pDawn(cI-LVA)-RBS070-X174E-T1 in response to blue photolysis in <i>G. hansenii</i> ATCC53582.</h3>
 
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<p>pDawn(cI-LVA)-RBS070-X174E-T1 was inserted into <i>G. hansenii</i>  ATCC53582 by electroporation and identified. As shown in Figure 4, pDawn(cI-LVA)-RBS070-X174E-T1 was successfully inserted into <i>G. hansenii</i> ATCC53582.</p>
 
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[[File:K27 5.png|600px|thumb|center|Figure4: Successfully identified by agarose gel electrophoresis (1)pSEVA331-pDawn(cI-LVA)-RBS070-LKD-T1. <i>G. hansenii</i> ATCC53582; (2)pSEVA331-pDawn(cI-LVA)-LKD-T1- <i> G.  hansenii</i> ATCC53582; (3)pSEVA331-pDawn(cI-LVA)-RBS070-X174E-T1- <i>G. hansenii</i> ATCC53582; (4)pSEVA331-pDawn(cI-LVA)-X174E-T1- <i>G. hansenii</i>  ATCC53582 and all of them were constructed successfully.]]
 
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<p>We transformed the pDawn (cI-LVA) -X174E-T1 plasmid into <i>G. hansenii</i> ATCC53582 and identified pDawn (cI-LVA) -X174E-T1 by using agarose gel electrophoresis. As shown in Figure 3, we successfully constructed the pDawn (cI-LVA) -X174E-T1.</p>
 
  
 
<h3>References</h3>
 
<h3>References</h3>
 
<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>
 
<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>

Revision as of 13:52, 11 October 2022


pDawn-RBS070-X174E-T1

Description

This composite part is a generator consisting of pDawn(cI-LVA)(BBa_K1075044) and X174E(BBa_K1835500).

Usage

The pDawn (cI-LVA)(BBa_K1075044) blue light response system and the lysis gene X174E(BBa_K1835500) were inserted into the pSEVA331 expression vector,which wsa inserted into the E.coli TOP10,screened out bacterial colony that grew in the dark but did not grow under blue light. Finally, the pSEVA331-pDawn-RBS070-X174E-T1 plasmid was selected and inserted into G. hansenii ATCC53582 to verify the responsiveness of pDawn(cI-LVA) to blue light.

Figure 1:Gene circuit of pDawn(cI-LVA)-RBS070-X174E-T1

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 pSEVA331-pDawn(cI-LVA)-RBS070-X174E-T1 in response to blue light lysis in E. coli.

As shown in Figure 2, the 11th and 15th bacterial colony did not grow under blue light but grow in dark. In order to further explore the lysis effect of pSEVA331-pDawn (cI-LVA)-RBS070-X174E-T1-TOP10, we measured the OD600 value of pSEVA331-pDawn (cI-LVA)-RBS070-X174E-T1-TOP10 periodically andploting the growth curve diagram.As shown in Figure 3, the lysis effect of pSEVA331-pDawn (cI-LVA)-RBS070-X174E-T1-TOP10 is not well.

Figure 2: The growth condition of pDawn(cI-LVA)-RBS070-X174E-T1-TOP10 in dark and under the blue light.
Figure 3: Growth curve diagram of pSEVA331-pDawn(cI-LVA)-RBS070-X174E-T1-TOP10.


References

[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.