Note: Please put up annotated/highlighted PDFs of papers in the 'Answers' pages
After General Wet Lab Meeting 1 , we came up with a set of questions to be answered.
They are as follows:
- What are the list of parts we will need for the planned genetic modifications?
- What are the corresponding BioBricks for these parts if any?
- What are the methods we will use to perform these modifications? (contact Prem and PhD mentors accordingly)
- What data can we get from Dry Lab that makes our life easier?
Please note that these questions have to be answered by all 3 groups
Questions specific to E. coli
- What strain of E. coli are we using?
- What is the final metabolic pathway expected to be after all engineering is done?
- Do we stick exactly to the Lin et al 2005 paper?
- How are knockouts done? (contact PhD mentors)
- What promoters are we going to put the genes under? (contact PhD mentors)
- What will be the final construct of the plasmid we will be using? (contact PhD mentors)
- How do we get the succinate out of the E. coli?
- How efficient should sucrose uptake be so as to not overwhelm the cyanobacteria?
- What is the final list of modifications that need to be made?
- What genes are we editing?
- glgc/invertase v/s sps/spp
- What are the final constructs we will be using for the transformation? (contact Prem)
Questions Specific to Cyanobacteria
- What strain of cyanobacteria are we using?
- What promoters are we going to put the genes under?
- What is the method of transformation that is most preferred for the strain we choose? (contact Prem)
Questions Specific to the co-culture
- How to make similar the growth rates of cyanobacteria and E. coli?
- Do we encapsulate the cyanobacteria and E. coli in the co-culture?
- Does this make assaying the growth rate easier?
- How do we get succinate out of the growth medium?
- What is the optimum salt stress & pH for the co-culture? + how will we maintain constancy in levels
- Sucrose production
- Succinate production
- Rate transfer assays for sucrose
- Expression assays (Wild Type v/s Mutant)