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NOTEBOOK

Here's a track of what we have done:

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  1. pYB1a-PobR-mCherry-Cmr constructing experiment
  2. pYT1a-ttgR-mCherry-Cmr constructing experiment
  3. pYB1a-mutator-T7(wt promoter) constructing experiment
  4. pRBlS-single-T7 constructing experiment
  5. pRBI2s-single-T7-STS constructing experiment
  6. pRl2s-reverse-T7 constructing experiment
  1. pYT1a-ttgR-mCherry-Kana constructing experiment
  2. pYB1a-mutator(mut) constructing experiment
  3. pLI2s-single or dual-T7-STS-oripir constructing experiment
  4. Mutation 1-8cycle
  1. pYT1a-ttgRmut-mCherry-Kana constructing experiment
  2. pYT1a-responsive-ttgR constructing experiment
  3. Mutation 9-21cycle
  4. Characterization based on these two plasmids pYT1a-ttgRmut-mCherry-Kana and pYT1a-responsive-ttgR
  1. glb2a-PpobA*2-mCherry-Cmr-SacB constructing experiment
  1. glb2a-PpobA*2-mCherry-Cmr-SacB constructing experiment
  2. gYb2a-PpobA*2-mCherry-SacB-Cmr constructing experiment
  3. Selection based on these two plasmids glb2a-PpobA*2-mCherry-Cmr-SacB and gYb2a-PpobA*2-mCherry-SacB-Cmr
  1. gYB2a-pobRWT-mCherry-codA-cmr constructing experiment
  2. Selection based on the plasmid gYB2a-pobRWT-mCherry-codA-cmr

Download : Here is our original notebook.

Here's a summary of what we have done.

Important experiments table
Date Member Experiment Result
7.20-8.27 Qianwen Jin, Fenglin Tao, Yuzhu Wang Constructed the plasmid gYB2a-pobRWT-mCherry-codA-cmr Our target plasmid has been constructed successfully
8.28-9.10 Xiaoya Wei, Yangbohui Yang, Jiale Li, Linshan Cao, Chengjie Dong Validation of PobRWT PobRWT exhibited a high degree of ligand specificity with a narrow dynamic range
Validating the feasibility of a dual selection system The dual selection system is feasible
Test the capacity of our mutant library The selection capacity for each compound was more than 900,000 clones (with at least four plates about 450,000 CFU/mL)
Dual selection significantly reduces the response of the sensor to natural substrates The obtained PobR mutants exhibited significantly reduced basal expression and robustly attenuated 4HB responsiveness compared to that of PobRWT
9.11-9.29 Jianing Li, Xinyao Yuan, Yuetong Zhu Selecting the PobR mutation library We obtained a biosensor that the fluorescence intensity was at least 1.5-fold higher than that of the negative control

Download : Here is detailed notebook mentioned on this table. If you want to see table stating which experiment contributed to which report figure, please click here.

Click here to see all our raw experimental data.

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