Difference between revisions of "Part:BBa K4325024"
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<h3>Characterization</h3> | <h3>Characterization</h3> | ||
<h4> | <h4> | ||
− | 1.Batch screening of pDawn( | + | 1.Batch screening of pDawn(cl)-X174E-T1 by evaluating the response to blue light.</h4> |
− | [[File:K24 1.png|600px|thumb|center|Figure1. <b>(1) pDawn(cI)-X174E-T1</b>; (2) pDawn(cI-LVA)-X174E-LVA-T1; (3) pDawn(cI)-LKD-T1; (4) pDawn(cI-LVA)-RBS070-LKD-LVA-T1 were constructed successfully.]]<p></p> [[File:K24 2.png|600px|thumb|center|Figure2. Growth condition of <i>E.coli </i> TOP10-pDawn(cI)-X174E-T1 in dark and under light.]] | + | [[File:K24 1.png|600px|thumb|center|Figure1. <b>(1) pDawn(cI)-X174E-T1</b>; (2) pDawn(cI-LVA)-X174E-LVA-T1; (3) pDawn(cI)-LKD-T1; (4) pDawn(cI-LVA)-RBS070-LKD-LVA-T1 were constructed successfully.]]<p></p> [[File:K24 2.png|600px|thumb|center|Figure2. Growth condition of <i>E.coli </i> TOP10-pDawn(cI)-X174E-T1 in the dark and under blue light.]] |
− | <p></p> As shown in Figure 1, we successfully constructed pDawn(cI)-X174E-T1 and transformed it into <i> E. coli</i> TOP10, | + | <p></p> As shown in Figure 1, we successfully constructed pDawn(cI)-X174E-T1 and transformed it into <i> E. coli</i> TOP10, selecting bacteria colonies and culturing in different growth conditions. As shown in Figure 2, the 9<sup>th</sup> colony of <i>E.coli</i> TOP10-pDawn(cI)-X174E-T1 grew in the dark, but did not grow under blue light. |
<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>[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. |
Latest revision as of 14:36, 12 October 2022
pDawn(cl)-X174E-T1
Description
This composite part is a generator consisting of pDawn(cI) (BBa_K4325010), X174E (BBa_K1835500) and T1 terminator (BBa_K3033016).
Usage
We inserted the blue light responsive system pDawn(cI) (BBa_K4325010) and the lysis gene X174E (BBa_K1835500) into the pSEVA331 expression vector, which was introduced into E. coli TOP10. The bacterial colonies that grew in the dark but did not grow under blue light were screened to verify the responsiveness of pDawn(cI) to blue light.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 2171
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE 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 - 1000INCOMPATIBLE 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)-X174E-T1 by evaluating the response to blue light.
As shown in Figure 1, we successfully constructed pDawn(cI)-X174E-T1 and transformed it into E. coli TOP10, selecting bacteria colonies and culturing in different growth conditions. As shown in Figure 2, the 9th colony of E.coli TOP10-pDawn(cI)-X174E-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] Witte, A., Wanner, G., Sulzner, M. & Lubitz, W. Dynamics of PhiX174 protein E-mediated lysis of Escherichia coli. Arch. Microbiol. 157, 381-8 (1992).