Difference between revisions of "Part:BBa K4325021"
 
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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-X174E-T1 by evaluating the response to blue light </h4>
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<h4>1.Batch screening of pDawn-X174E-T1 by evaluating the response to blue light. </h4>
 
[[File:K21 2.png|600px|thumb|center|Figure 2: Growth condition of <i>E. coli</i> TOP10-pDawn(cI-LVA)-X174E-T1-TOP10 in the dark and under light.]]<p></p>
 
[[File:K21 2.png|600px|thumb|center|Figure 2: Growth condition of <i>E. coli</i> TOP10-pDawn(cI-LVA)-X174E-T1-TOP10 in the dark and under light.]]<p></p>
 
[[File:K21 3.png|600px|thumb|center|Figure 3: Growth curve diagram  of <i> E. coli</i> TOP10-pDawn(cI-LVA)-X174E-T1-TOP10.]]<p></p>
 
[[File:K21 3.png|600px|thumb|center|Figure 3: Growth curve diagram  of <i> E. coli</i> TOP10-pDawn(cI-LVA)-X174E-T1-TOP10.]]<p></p>

Latest revision as of 14:34, 12 October 2022

pDawn-X174E-T1

Description

This composite part is a generator consisting of pDawn(cI-LVA) (BBa_K1075044) ,X174E (BBa_K1835500) and T1 terminator (BBa_K3033016).

Figure 1: Genetic circuit of pDawn(cI-LVA)-X174E-T1.

Usage

We inserted the blue light responsive system pDawn(cI-LVA) (BBa_K1075044) and the lysis gene X174E (BBa_K1835500) into the pSEVA331 expression vector, which was introduced into E. coli TOP10 and screened for colonies that grew in the dark but did not grow under 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-T1 by evaluating the response to blue light.

Figure 2: Growth condition of E. coli TOP10-pDawn(cI-LVA)-X174E-T1-TOP10 in the dark and under light.

Figure 3: Growth curve diagram of E. coli TOP10-pDawn(cI-LVA)-X174E-T1-TOP10.

As shown in Figure 2, the 1th , 4th , 5th, 6th, 7th, 8th, 10th, 14th, and 16th colonies met our expectations. To further search the lysis effect of pDawn(cI-LVA) -X174E-T1, we tracked the OD600 value of E. coli TOP10-pSEVA311-pDawn(cI-LVA)-X174E-T1 and plotted the growth curves diagram as shown in Figure 3. It shows that the lysis effect of pDawn(cI-LVA)-X174E-T1 is mediocre.

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).