Difference between revisions of "Part:BBa K4325026"
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<partinfo>BBa_K4325026 short</partinfo> | <partinfo>BBa_K4325026 short</partinfo> | ||
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===Usage=== | ===Usage=== | ||
<p>The pDawn (cI-LVA) blue light response system (<partinfo>BBa_K1075044</partinfo>) and lysis gene LKD (<partinfo>BBa_K4325004</partinfo>) were inserted into the pSEVA331 expression vector, which was inserted into <i>E. coli</i> TOP10, screen out the bacterial colony that grew in the dark but did not grow under blue light Finally, the pSEVA331-pDawn-RBS070-LKD-T1 plasmid was extracted and inserted by electroporation into <i> G. hansenii</i> ATCC53582 to verify the responsiveness of pDawn (cI-LVA) to blue light.</p> | <p>The pDawn (cI-LVA) blue light response system (<partinfo>BBa_K1075044</partinfo>) and lysis gene LKD (<partinfo>BBa_K4325004</partinfo>) were inserted into the pSEVA331 expression vector, which was inserted into <i>E. coli</i> TOP10, screen out the bacterial colony that grew in the dark but did not grow under blue light Finally, the pSEVA331-pDawn-RBS070-LKD-T1 plasmid was extracted and inserted by electroporation into <i> G. hansenii</i> ATCC53582 to verify the responsiveness of pDawn (cI-LVA) to blue light.</p> | ||
| − | [[File:K26 | + | [[File:K26 11.png|600px|thumb|center|Figure 1: <p></p>Gene circuit of pDawn (cI-LVA)-RBS070-LKD-T1.]] |
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<p>As shown in Figure 2,except for the 11th and 13<sup>th </sup> bacterial colony,almost all of the bacterial colony did not grow under blue light.To further explore the lysis effect of pSEVA331-pDawn (cI-LVA) -RBS070-LKD-T1-TOP10, we measured the OD <sub>600</sub> values of pSEVA331-pDawn (cI-LVA) -RBS070-LKD-T1-TOP10 and ploting the growth curve diagram.As shown in Figure 3, the pSEVA331-pDawn (cI-LVA) -RBS070-LKD-T1-TOP10 lysis effect was well in the first eight hours, but was unstable after fifteen hours.</p> | <p>As shown in Figure 2,except for the 11th and 13<sup>th </sup> bacterial colony,almost all of the bacterial colony did not grow under blue light.To further explore the lysis effect of pSEVA331-pDawn (cI-LVA) -RBS070-LKD-T1-TOP10, we measured the OD <sub>600</sub> values of pSEVA331-pDawn (cI-LVA) -RBS070-LKD-T1-TOP10 and ploting the growth curve diagram.As shown in Figure 3, the pSEVA331-pDawn (cI-LVA) -RBS070-LKD-T1-TOP10 lysis effect was well in the first eight hours, but was unstable after fifteen hours.</p> | ||
| − | [[File:K26 | + | [[File:K26 22.png|600px|thumb|center|Figure 2: <p></p>Growth condition of pDawn (cI-LVA)-RBS070-LKD-T1-TOP10 in the dark and under the light]] |
[[File:K26 3.png|600px|thumb|center|Figure 3: <p></p> Growth curve diagramof pDawn (cI-LVA)-RBS070-LKD-T1-TOP10]] | [[File:K26 3.png|600px|thumb|center|Figure 3: <p></p> Growth curve diagramof pDawn (cI-LVA)-RBS070-LKD-T1-TOP10]] | ||
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<p>As shown in Figure 3, we successfully indicated the pDawn (cI-LVA) -RBS070-LKD-T1 into <i>G. hansenii</i> ATCC53582 by electroporation</p> | <p>As shown in Figure 3, we successfully indicated the pDawn (cI-LVA) -RBS070-LKD-T1 into <i>G. hansenii</i> ATCC53582 by electroporation</p> | ||
| − | [[File:K26 | + | [[File:K26 44.png|600px|thumb|center|Figure 4: <p></p>Successfully identification withagarose gel electrophoresis <p></p> |
(1)pSEVA331-pDawn(cI-LVA)-RBS070-LKD-T1-<i>G. hansenii</i> ATCC53582 <p></p> | (1)pSEVA331-pDawn(cI-LVA)-RBS070-LKD-T1-<i>G. hansenii</i> ATCC53582 <p></p> | ||
(2)pSEVA331-pDawn (cI-LVA)-LKD-T1-<i>G. hansenii</i> ATCC53582 <p></p> | (2)pSEVA331-pDawn (cI-LVA)-LKD-T1-<i>G. hansenii</i> ATCC53582 <p></p> | ||
Revision as of 10:26, 10 October 2022
pDawn-RBS070-LKD-T1
Description
This composite part is a generator containing pDawn (cI-LVA) (BBa_K1075044) and LKD (BBa_K4325004).
Usage
The pDawn (cI-LVA) blue light response system (BBa_K1075044) and lysis gene LKD (BBa_K4325004) were inserted into the pSEVA331 expression vector, which was inserted into E. coli TOP10, screen out the bacterial colony that grew in the dark but did not grow under blue light Finally, the pSEVA331-pDawn-RBS070-LKD-T1 plasmid was extracted and inserted by electroporation into G. hansenii ATCC53582 to verify the responsiveness of pDawn (cI-LVA) 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 pSEVA331-pDawn(cI-LVA)-RBS070-LKD-T1 in response to blue light lysis in E. coli TOP10.
As shown in Figure 2,except for the 11th and 13th bacterial colony,almost all of the bacterial colony did not grow under blue light.To further explore the lysis effect of pSEVA331-pDawn (cI-LVA) -RBS070-LKD-T1-TOP10, we measured the OD 600 values of pSEVA331-pDawn (cI-LVA) -RBS070-LKD-T1-TOP10 and ploting the growth curve diagram.As shown in Figure 3, the pSEVA331-pDawn (cI-LVA) -RBS070-LKD-T1-TOP10 lysis effect was well in the first eight hours, but was unstable after fifteen hours.
pSEVA331-pDawn(cI-LVA)-RBS070-LKD-T1 in response to blue photolysis in G. hansenii ATCC53582.
As shown in Figure 3, we successfully indicated the pDawn (cI-LVA) -RBS070-LKD-T1 into G. hansenii ATCC53582 by electroporation
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.