Team:Bielefeld-Germany/Results/Characterization
From 2010.igem.org
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===<partinfo>P1010</partinfo>: ''ccdB''-gene=== | ===<partinfo>P1010</partinfo>: ''ccdB''-gene=== | ||
- | We transformed this BioBrick into ''Escherichia coli'' JM109, DH5α, TOP10 and DB3.1. ''E. coli'' JM109 and DH5α seem to be ''ccdB'' resistant because there were as much colonies after P1010 transformation as observed with DB3.1. The P1010 works as expected in ''E. coli'' TOP10 (no colonies after transformation) and DB3.1 (lawn after transformation). | + | The ''ccdB'' gene targets the gyrase of ''E. coli'' and is lethal for all ''E. coli'' strains without the gyrase mutation gyrA462. The ''ccdB'' BioBrick is used for the 3A-assembly as a positive selection marker. |
+ | We transformed this BioBrick into ''Escherichia coli'' JM109, DH5α, TOP10, XL1-Blue, EC100D and DB3.1. ''E. coli'' JM109 and DH5α seem to be ''ccdB'' resistant because there were as much colonies after P1010 transformation as observed with DB3.1. The P1010 works as expected in ''E. coli'' TOP10 (no colonies after transformation) and DB3.1 (lawn after transformation). | ||
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===<partinfo>K389011</partinfo>: VirA screening device=== | ===<partinfo>K389011</partinfo>: VirA screening device=== |
Revision as of 13:42, 10 October 2010
Contents |
Tested BioBricks
We tested the following BioBricks:
<partinfo>K238008</partinfo>: virA
We made a restriction analysis and sequenced parts of this BioBrick. There were problems - more information within a short time.
<partinfo>BBa_K238011</partinfo>: vir-promoter
We made a restriction analysis and sequenced parts of this BioBrick.
<partinfo>P1010</partinfo>: ccdB-gene
The ccdB gene targets the gyrase of E. coli and is lethal for all E. coli strains without the gyrase mutation gyrA462. The ccdB BioBrick is used for the 3A-assembly as a positive selection marker. We transformed this BioBrick into Escherichia coli JM109, DH5α, TOP10, XL1-Blue, EC100D and DB3.1. E. coli JM109 and DH5α seem to be ccdB resistant because there were as much colonies after P1010 transformation as observed with DB3.1. The P1010 works as expected in E. coli TOP10 (no colonies after transformation) and DB3.1 (lawn after transformation).
Table 1: Results of the transformation of the cell-death gene ccdB, BioBrick <partinfo>P1010</partinfo>, into different strains of E. coli.
E. coli strain | Resistant to ccdB? | Expected result? | Gyrase genotype |
---|---|---|---|
DB3.1 | yes | yes | gyrA462 |
DH5α | yes | no | gyrA96 |
JM109 | yes | no | gyrA96 |
TOP10 | no | yes | WT |
<partinfo>K389011</partinfo>: VirA screening device
![](/wiki/images/f/fe/IGEM-Bielefeld-LD50.jpg)
Sequenced BioBricks
Own BioBricks
The sequencing of the following of our own BioBricks was succesful and lead to the expected results:
- <partinfo>K389001</partinfo> (not fully completed)
- <partinfo>K389002</partinfo>
- <partinfo>K389003</partinfo>
- <partinfo>K389004</partinfo>
- <partinfo>K389005</partinfo>
- <partinfo>K389010</partinfo> (not fully completed)
- <partinfo>K389011</partinfo> (not fully completed)
- <partinfo>K389012</partinfo> (not fully completed)
- <partinfo>K389013</partinfo> (not fully completed)
- <partinfo>K389015</partinfo> (not fully completed)
- <partinfo>K389016</partinfo> (not fully completed)
- <partinfo>K389050</partinfo>
Other BioBricks
- <partinfo>K238008</partinfo> sequencing gave negative results - infos in registry are not correct!
- <partinfo>K238011</partinfo> sequencing gave negative results - infos in registry are not correct!