March
3/4
- Topic decision
- Information collection
3/11
- 7 chemical decided: Astaxanthin, NMN, PQQ, Vit B6, Butyrate, HMB(β-Hydroxy β-methylbutyric acid), KIC(α-Ketoisocaproic acid)
- The seven chemicals’ effects on human
- mTOR signal transduction pathway research
3/18
- Market analyzing of the Sarcopenia
- Data collection on Sarcopenia
- Research on supportive treatment of Sarcopenia
3/25
- Research on the cause of the Sarcopenia
- Research on the mechanism of Butyrate and HMB to boost muscle production
- Deeper research in mTOR pathway
April
4/1
- The connection of mTOR and Butyrate and HMB
- Research the mechanism of boosting muscle of other compounds:Astaxanthin, NMN, PQQ, Vit B6,and KIC
- Creating 31 pages report for our research
![](https://static.igem.wiki/teams/4743/wiki/notebook-3.png)
![](https://static.igem.wiki/teams/4743/wiki/notebook-4.png)
4/8
- The connection of mTOR and Butyrate and HMB
- Research the mechanism of boosting muscle of other compounds:Astaxanthin, NMN, PQQ, Vit B6,and KIC
- Creating a 31 page report for our research on 7 chemical compounds
4/15
- Decide the organism of producing these compounds
- Recruit new members
- Creating introductory presentation on PPT
- Done with our drawing of chemical compounds
![](https://static.igem.wiki/teams/4743/wiki/notebook-6.png)
4/22
- Distribute the works of producing chemical compounds to three probiotics
1.Clostridium Butyricum: Butyrate, Vitamin B6
2.Bacillus subtilis: PQQ, HMB, KIC
3.Kluyveromyces marxianus: NMN, Astaxanthin
- Looking for metabolic pathways for producing these compounds on KEGG or other resource
- Done the First draft on metabolic pathway producing NMN
4/29
- Decide the knockouts of the genes
- Starting to search for possible kill switch
![](https://static.igem.wiki/teams/4743/wiki/notebook-9.png)
May
5/13
- Decide to concentrate on NMN,HMB,and KIC production
- Done the second draft of NMN producing pathway
5/20
- Done the final draft of NMN producing pathway
- Designed our own pathway for NMN production
![](https://static.igem.wiki/teams/4743/wiki/notebook-12.png)
5/27
- Interview with doctors
- Kill switch design by using operon system
- Learned on biobrick build up
June
6/3
- Research on the conformation of the enzyme NAMPT(an enzyme essential for NMN)
- BLAST and search for the highest efficiency of NAMPT to produce most NMN
- Finalize the pathway to produce HMB and KIC
6/10
- Keep researching on NAMPT
- Researching for the Intestine environment and the location of our probiotics will grow
- Researching for dosage of our chemical compounds
![](https://static.igem.wiki/teams/4743/wiki/notebook-15.png)
6/17
- Finding out sequence for what we want to insert and what we want to knock out in our metabolic pathway
- Design the second draft of our kill switch for Bacillus Subtilis
6/24
- Finding out key active site of the NAMPT
- Design the third draft of our kill switch for Bacillus Subtilis
- Research for hydrogen sulfide sensitive repressor
- Finding Toxin and Antitoxin of Bacillus Subtilis
![](https://static.igem.wiki/teams/4743/wiki/notebook-17.png)
July
7/1
- Draw out the kill switch for Bacillus Subtilis
- Drawing the first draft of kill switch of Clostridium and kluyveromyces
7/8
- Choosing the expression vector of Bacillus subtilis
- Preparing for competent cells
- Draw out the gene map
7/15
- Learning to use snapgene to draw map
- Design the map of kluyveromyces marxianus
![](https://static.igem.wiki/teams/4743/wiki/notebook-21.png)
7/22
- Send out the order of fragments
- Sequence optimization
- Optimize the design
7/29
- Finalize the genes and order out
- Complete the transformation of TDH3,ADH1 promoter and GFP gene from the distribution kit
- Done the colon PCR
August
8/5
8/12
- Start the experiments of NadA,NadB,and NadE fragments
- Ligation and transformation for the plasmid
- Gel extraction
- Check the length of pKlac2 plasmid
![](https://static.igem.wiki/teams/4743/wiki/notebook-23.png)
8/19
- Colony PCR checking
- Failed the ligation
8/26
- Started Astaxanthin experiment
- Planning to redo the NMN experiment
September
9/2
- Wiki build up- HP and wet lab
- Final presentation making
9/9
- Wiki build up- Projet,wet lab, and Human practice
- Enter register
- Started making promotion videos
9/16
- Wiki build up- Projet,wet lab, and Human practice
- Checking for safety forms
- Finalize promotion videos
9/23
- Wiki build up- Projet,wet lab, and Human practice
- Finalizing safety forms
- Finalize promotion videos