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Team:EPF-Lausanne/Our Project/TetR mutants/muTetRs
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==How== | ==How== | ||
| - | First we chose the <html> <a href="https://2011.igem.org/Team:EPF-Lausanne/Protocols/Site-specific_mutagenesis"> Site-specific mutagenesis </a></html> approach to make the mutants of TetR but that strategy <html> <a href="https://2011.igem.org/Team:EPF-Lausanne/Notebook/July2011#Tuesday.2C_12_July_2011"> was not working well </a> </html> and another one was tried in parallel: | + | First we chose the <html> <a href="https://2011.igem.org/Team:EPF-Lausanne/Protocols/Site-specific_mutagenesis"> Site-specific mutagenesis </a></html> approach to make the mutants of TetR but that strategy <html> <a href="https://2011.igem.org/Team:EPF-Lausanne/Notebook/July2011#Tuesday.2C_12_July_2011"> was not working well </a> </html> and another one was tried in parallel: <html> <a href="https://2011.igem.org/Team:EPF-Lausanne/Protocols/TetR_Extension_PCR" PCR-induced mutagenesis.</a></html> |
Site-directed mutagenesis consists of amplifying plasmid and | Site-directed mutagenesis consists of amplifying plasmid and | ||
Revision as of 01:00, 22 September 2011
Mutant TetRs
In vitro Main | Why TetR? | Mutant TetRs | MITOMI Data | In-vivo & In-vitro outlineHow
First we chose the Site-specific mutagenesis approach to make the mutants of TetR but that strategy was not working well and another one was tried in parallel:
Site-directed mutagenesis consists of amplifying plasmid and
What
| TetR variants | Mutagenesis | Promoter | Cterminal | DNA form | Expression | MITOMI vs 1-off | Biobrick | Sequenced | Sent to registry |
|---|---|---|---|---|---|---|---|---|---|
| V36F | site directed | SP6 | eGFP | plasmid | worked well | X | BBa_K613013 | ||
| V36FW43S | PCR-induced | T7 | His-tag | linear | (ins C-term), ITT tested | BBa_K613014 | |||
| E37AW43ST141A | PCR-induced | T7 | His-tag | linear | ITT tested | X | BBa_K613015 | X | X |
| P39K | PCR-induced | T7 | His-tag | template | ITT tested, worked | X | BBa_K613016 | X | X |
| Y42F | PCR-induced | T7 | His-tag | linear | ITT tested | X | BBa_K613017 | X | X |
| Y42FK108E | PCR-induced | T7 | His-tag | linear | ITT tested | BBa_K613018 | X | X | |
| P39QY42M | site directed | SP6 | eGFP | plasmid | worked well | X | BBa_K613019 | ||
| P39QY42ML197S | PCR-induced | T7 | His-tag | linear | ITT tested | BBa_K613020 | |||
| P39QY42ML52P | PCR-induced | T7 | His-tag | linear | ITT tested | BBa_K613021 | |||
| E37AP39K | site directed | T7 | His-tag | linear | BBa_K613022 | X | |||
| E37AP39QY42F | site directed | T7 | His-tag | linear | BBa_K613023 | X | |||
| WT | none | T7 | His&GFP | both |
