Meeting with Prem
| Date | |
|---|---|
| Department | GeneralHP |
| Description | Wangikar's PhD student |
| HP sub-branch | |
| Links/media | |
| Participants | |
| Property | |
| Property 1 | |
| Property 2 | |
| Type | Meetings |
- Decide on which strain to work with - PCC 7942, PCC 7002, UTEX 2973 - their lab has all of these strains?
- Synechocystis - if we choose to work with this we have to first look at availability becuase their lab may not have this
- Their lab isolated two fast growing strains - PCC 11801 and PCC 11802, 11801 grows faster than 7942? - doubling time of 2 days compared to 4 days for 7942
- Find out the optimal growing conditions - alkalinity/pH and the cloning techniques - suicide vectors, neutral sites, gibson assembly. 2973 can't be transformed naturally
- Co-culture should be feasible, salt effect on E coli shouldn't be too much of an issue, cyanobacteria doesnt need so much salt that it wont allow E coli to grow, contamination of cyanobacteria cultures with E coli in their lab is common
- Things needed for cyanobacteria culture - a decent shaking incubator with light source, proper media composition
- Recommended checking out 'CyanoGate'
http://www.plantphysiol.org/content/180/1/39
https://www.addgene.org/kits/mccormick-cyanogate/
- Someone in our lab just started working on sucrose production in cyanobacteria and they also ordered all the reagents for it, it should be possible for Wangikar to host you so please do ask him. You could do cyanobacteria work at our lab in parallel to e coli work in your institute.
- Ensure your pitch to wangikar is solid and you make a good impression, he'll want to know all details about the products you want to synthesise, their yields and prices.
- DECIDE ON A FINAL PRODUCT!!! Make sure it is a cool product, and try to show that your project is benefiting the environment and lowering costs. Compare prices and yields for whatever you're synthesising in E coli and Cyanobacteria
- Make your project simple with not more than 2-3 gene modifications in E coli. Keep engineering in cyanobacteria minimum because it will take a lot more time than E coli. Just do overexpression of sucrose-producing genes and exporting of sucrose in cyanobacteria. Time for cloning Cyanobacteria won't change whether 2 people are doing or if 10 are doing it. You'll still have to wait the same amount of time.
- Someone had just presented 2020 3-HP paper in Wangikar's lab.
- Even if media needs to contain some amount of glucose, compare how much glucose reduction you have achieved in medium by using sucrose secreting cyanobacteria.
- Look at how sucrose will be used by the cyanobacteria, will it use it as just as energy source or will it convert into something else?
- She'll provide details of media - possible BG-11 with 3% NaCl, will also send Damini Jaiswal papers.