Difference between revisions of "Part:BBa K1223007:Design"
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===Design Notes=== | ===Design Notes=== | ||
| − | This part is an improvement of the existing BBa_K327018 LVA tagged cI repressor protein. We added a his tag instead of the LVA tag in order to offer a convenient way to study the protein without damaging it. The his tag is located at its C-terminal, therefore having no effect on its function | + | This part is an improvement of the existing BBa_K327018 LVA tagged cI repressor protein. We added a his tag instead of the LVA tag in order to offer a convenient way to study the protein without damaging it. The his tag is located at its C-terminal, therefore having no effect on its function. |
| + | |||
| + | we used this part in order to identify our protein through western blot technique, using anti His-Tag antibodies. | ||
| + | in the western it is apparent that without induction there is no leakage of the cI protein but upon induction we get | ||
| + | a nice band indicating that the cI is expressed. The positive control we used was a model protein containing a His tag. | ||
| + | |||
| + | [[File:cIwestern.jpg]] | ||
| + | |||
| + | and coomassie staining of the gel. | ||
| + | |||
| + | [[File:westcom.jpg]] | ||
===Source=== | ===Source=== | ||
| − | synthetic. | + | part is synthetic. the cI CDS (Originated from the lambda bacteriophage) and the promoter were taken from the iGEM registry. |
===References=== | ===References=== | ||
| + | M. Pedersen, M. Ligowska, K. Hammer, Characterization of the CI repressor protein encoded by the temperate lactococcal phage, Journal of Bacteriology 29, [2010] | ||
Latest revision as of 13:30, 12 October 2013
cI translational unit with His-tag
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
Design Notes
This part is an improvement of the existing BBa_K327018 LVA tagged cI repressor protein. We added a his tag instead of the LVA tag in order to offer a convenient way to study the protein without damaging it. The his tag is located at its C-terminal, therefore having no effect on its function.
we used this part in order to identify our protein through western blot technique, using anti His-Tag antibodies. in the western it is apparent that without induction there is no leakage of the cI protein but upon induction we get a nice band indicating that the cI is expressed. The positive control we used was a model protein containing a His tag.
and coomassie staining of the gel.
Source
part is synthetic. the cI CDS (Originated from the lambda bacteriophage) and the promoter were taken from the iGEM registry.
References
M. Pedersen, M. Ligowska, K. Hammer, Characterization of the CI repressor protein encoded by the temperate lactococcal phage, Journal of Bacteriology 29, [2010]