Difference between revisions of "Part:BBa K1223007"
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<partinfo>BBa_K1223007 short</partinfo> | <partinfo>BBa_K1223007 short</partinfo> | ||
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lambda cI translational unit with AraC/Lac promoter (BBa_K354000) and C-term His tag | lambda cI translational unit with AraC/Lac promoter (BBa_K354000) and C-term His tag | ||
| − | + | ===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. | ||
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| + | 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. | ||
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| + | [[File:cIwestern.jpg]] | ||
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| + | and coomassie staining of the gel. | ||
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| + | [[File:westcom.jpg]] | ||
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| + | ===Source=== | ||
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| + | 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] | ||
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===Usage and Biology=== | ===Usage and Biology=== | ||
Latest revision as of 13:31, 12 October 2013
cI translational unit with His-tag
lambda cI translational unit with AraC/Lac promoter (BBa_K354000) and C-term His tag
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]
Usage and Biology
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]