Team:LMSU/Goals

# Our goals

### Assemble all the necessary constructions using Golden gate protocol

During the design stage of our experiment, we decided to assemble the structure using the golden gate method. We were able to assemble a number of structures, however, we encountered insufficient effectiveness of the methodology we used. In order to use Golden Gate as the main method of assembling structures in the future, one of our top priorities for the next year will be troubleshooting. We plan to consult with experts in molecular cloning techniques to optimize the assembly method for the constructs required for this project.

### Comparison of the strength of promoters and RBS in Escherichia coli and Arthrospira platensis

We organized a collaboration with colleagues from team: Madrid in order to further extrapolate our results in an experiment where we tested the strength of promoters in *E. coli* and *Arthrospira platensis*. However, we have received the results of determining the strength of promoters and their inducibility in *E. coli*, and are ready for the next step in new research.

### Whole genome sequencing of Arthrospira platensis and selection of locus for transformation

Since we plan to insert the construct into the genome using site-specific recombination, it is necessary to carry out whole genome sequencing of the selected *Arthrospira platensis* strain IPPAS B-256. Obtaining the sequence of the entire genome will make it possible to better select the optimal loci for site-specific recombination in the strain of our choice.

### Selection of optimal conditions for the transformation of Arthrospira platensis

The transformation of Arthrospira platensis, like many other cyanobacteria, is a non-trivial task. Due to the fact that the conditions need to be selected for a long time and scrupulously, we focused our efforts on tasks that are easier to implement. However, our work cannot exist without a stage of transformation.

### Galactosidase repression test

We plan to test the efficiency of the repression of the promoter by the GAL4 repressor. For this, a system will be made, where galactosidase will act as a reporter, and its presence in the cell will be detected by a standard color reaction. If the repression is carried out effectively, then this will be noticeable by the result of galactosidase activity.

### Modeling of RNA-ribosome stoichiometry to increase the strength of RBS-sequences

The optimal choice of RBSa is one of the main tools for the expression of the gene of interest. Therefore, we plan to simulate the interaction of 16s-rRNA with different RBSs for efficient efficient RBSa.

### Cultivation of Arthrospira platensis on the medium, which is urine as the main source

One of the key tasks of our project is the cultivation of an arthrospira on the crew's waste. Urea can act as a nitrogen source for arthrospira (Heisoo, D., 2014). Also, with its growth, exhaled CO2 is utilized. The maximum use of waste products is a necessity for long space travels. We recommend our selection of the optimal medium for growing *Arthrospira platensis*.