Difference between revisions of "Part:BBa K4325025"
 
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<partinfo>BBa_K4325025 short</partinfo>
 
<partinfo>BBa_K4325025 short</partinfo>
 
===Description===
 
===Description===
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The composite part is a generator consisting of pDawn (without LVA tag) (<partinfo>BBa_K4325010 </partinfo>) and LKD (<partinfo>BBa_K4325004 </partinfo>).
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The composite part is a generator consisting of pDawn(cI)(<partinfo>BBa_K4325010 </partinfo>), RBS070 (<partinfo>BBa_K4325002 </partinfo>), LKD(<partinfo>BBa_K4325004 </partinfo>) and T1 terminator (<partinfo>BBa_K3033016 </partinfo>).
 
===Usage===
 
===Usage===
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<p>We inserted the pDawn (without LVA tag) (<partinfo>BBa_K4325010 </partinfo>)blue light response system and the lysis gene LKD (<partinfo>BBa_K4325004</partinfo>) into the pSEVA331 expression vector, which was inserted into <i> E. coli</i> TOP10 screened out the colonies grew in the dark but did not grow in blue light to verify the responsiveness of pDawn (without LVA tag) to blue light.</p >
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<p>We introduced the blue light responsive system pDawn(cI) (<partinfo>BBa_K4325010 </partinfo>) and the lysis gene LKD (<partinfo>BBa_K4325004 </partinfo>) into the pSEVA331 expression vector, which was incorporated into <i>E. coli</i> TOP10 and screened for the colonies that grew in the dark but did not grow under blue light to verify the responsiveness of pDawn(cI) to blue light.</p >
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===Sequence and Features===
 
===Sequence and Features===
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<h3>Characterization</h3>
 
<h3>Characterization</h3>
 
<h4>
 
<h4>
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1.Batch screening of pDawn(without LVA tag)-RBS070-LKD-T1-pSEVA331 in response to blue light lysis in <i> E. coli</i>.</h4>
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1.Batch screening of pDawn(cl)-RBS070-LKD-T1 by evaluating the response to blue light.</h4>
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[[File:K025 without LVA.png|600px|thumb|center|Figure 1 was the growth condition of pSEVA331-pDawn (without LVA tag) -RBS070-LKD-T1-TOP10 both in the dark and under the light.]]<P></P>As shown in Figure 1, four bacterial colony of pSEVA331-pDawn (without LVA tag) -RBS070-LKD-T1-TOP10 grew in the dark, but it did not grow under the light.
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[[File:K025 without LVA.png|600px|thumb|center|Figure 1:  Growth condition of <i>E.coli</i> TOP10-pSEVA331-pDawn(cI)-RBS070-LKD-T1 in the dark and under blue light.]]<P></P>
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As shown in Figure 1, 10 bacterial colonies of <i>E.coli</i> TOP10-pSEVA331-pDawn(cI)-RBS070-LKD-T1 grew in the dark, but did not grow under blue light.
 
<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] 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>[2] [2]Ceyssens, P.-J. et al. Genomic analysis of Pseudomonas aeruginosa phages LKD16 and LKA1: establishment of the phiKMV subgroup within the T7 supergroup. J. Bacteriol. 188, 6924-31 (2006).</p >

Latest revision as of 14:40, 12 October 2022


pDawn(cl)-RBS070-LKD-T1

Description

The composite part is a generator consisting of pDawn(cI)(BBa_K4325010), RBS070 (BBa_K4325002), LKD(BBa_K4325004) and T1 terminator (BBa_K3033016).

Usage

We introduced the blue light responsive system pDawn(cI) (BBa_K4325010) and the lysis gene LKD (BBa_K4325004) into the pSEVA331 expression vector, which was incorporated into E. coli TOP10 and screened for the colonies that grew in the dark but did not grow under blue light to verify the responsiveness of pDawn(cI) 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(cl)-RBS070-LKD-T1 by evaluating the response to blue light.

Figure 1: Growth condition of E.coli TOP10-pSEVA331-pDawn(cI)-RBS070-LKD-T1 in the dark and under blue light.

As shown in Figure 1, 10 bacterial colonies of E.coli TOP10-pSEVA331-pDawn(cI)-RBS070-LKD-T1 grew in the dark, but did not grow under blue light.

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.

[2] [2]Ceyssens, P.-J. et al. Genomic analysis of Pseudomonas aeruginosa phages LKD16 and LKA1: establishment of the phiKMV subgroup within the T7 supergroup. J. Bacteriol. 188, 6924-31 (2006).