Difference between revisions of "Part:BBa K3408011"
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This composite part consists of promoter P<sub>liaG</sub> and P<sub>grac</sub>, lacI gene, CⅠ gene, GFP gene and some essential RBS and terminators. Promoter P<sub>liaG</sub> starts transcription process and its transcription production is LacI, which is regulated by IPTG. When bacteria is exposed to medium which is rich in IPTG, bacteria can intake IPTG. IPTG is combined with LacI, then LacI cannot be bound to promoter Pgrac which is specially suppressed by LacI. Therefore, CⅠ repressor protein cannot generate as downstream cⅠ gene cannot transcribe regularly. | This composite part consists of promoter P<sub>liaG</sub> and P<sub>grac</sub>, lacI gene, CⅠ gene, GFP gene and some essential RBS and terminators. Promoter P<sub>liaG</sub> starts transcription process and its transcription production is LacI, which is regulated by IPTG. When bacteria is exposed to medium which is rich in IPTG, bacteria can intake IPTG. IPTG is combined with LacI, then LacI cannot be bound to promoter Pgrac which is specially suppressed by LacI. Therefore, CⅠ repressor protein cannot generate as downstream cⅠ gene cannot transcribe regularly. | ||
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As a result, P<sub>CⅠ</sub> recovers its activity and triggers the downstream transcription of GFP. By checking the expression of GFP, we could verify the feasibility of IPTG induction system, thus guaranteeing the successful culture of our engineered <i>Bacillus subtilis</i>. | As a result, P<sub>CⅠ</sub> recovers its activity and triggers the downstream transcription of GFP. By checking the expression of GFP, we could verify the feasibility of IPTG induction system, thus guaranteeing the successful culture of our engineered <i>Bacillus subtilis</i>. | ||
Revision as of 14:33, 16 October 2020
PliaG-B0034-lacI-Pgrac-B0034-CⅠ-PCⅠ-GFP-B0015
This composite part consists of promoter PliaG and Pgrac, lacI gene, CⅠ gene, GFP gene and some essential RBS and terminators. Promoter PliaG starts transcription process and its transcription production is LacI, which is regulated by IPTG. When bacteria is exposed to medium which is rich in IPTG, bacteria can intake IPTG. IPTG is combined with LacI, then LacI cannot be bound to promoter Pgrac which is specially suppressed by LacI. Therefore, CⅠ repressor protein cannot generate as downstream cⅠ gene cannot transcribe regularly.
As a result, PCⅠ recovers its activity and triggers the downstream transcription of GFP. By checking the expression of GFP, we could verify the feasibility of IPTG induction system, thus guaranteeing the successful culture of our engineered Bacillus subtilis.
Plasmid profile
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 1294
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI.rc site found at 2155