The main project is the facile purification of DNA manipulating enzymes through fusing such enzymes to elastin-like polypeptides (ELPs). ELPs undergo a reversible phase transition in response to NaCl concentrations, in that raising the NaCl concentration of an ELP solution will cause the ELPs to aggregate. These aggregates can easily be separated from a complex mixture via centrifugation or microfiltration, and re-solubilized by lowering the NaCl concentration; a process termed ‘inverse transition cycling’ (ITC). Addition of a responsive self-cleaving peptide in between the protein of interest and the ELP, followed by another round of ITC, results in the purification of functional proteins.
We have two goals for this project:
1. To improve the ITC method by addition of a secretion tag to the ELP fusion proteins, precluding the need for cell lysis, as well as enabling the possibility of an efficient continuous production system.
2. Automation of the entire process through collaboration with computer scientists and roboticists which share our community maker-space.
The vision of our final product is a robot that produces functional workhorse enzymes on the bench top. People can create novel constructs and upload the sequence online, where others can download it and create in-house cell lines for the enzymes they need, which would all be purified by the robot. This would lower the burden of cost for lab work, lowering the bar of entry for entrepreneurs and innovators in the synthetic biology field.
A small aquarium pump, some tubing, plastic bottles and hot glue are all that’s needed to make a photobioreactor. Our set-up is on display because it’s green, it bubbles and it looks cool as well as to show that scientific experiment is not an exotic affair, it’s within reach.
Experiment 1: Unprocessed Algae as fuel source
Step 1: Dehydrate Algae.
Step 2: Burn it
The current community project is species typing in commercial meat. DNA is extracted from the meat sample and species-specific primers are used to amplify DNA in the sample via the polymerase-chain-reaction (PCR). Analysis of the resulting DNA fragments shows which animal species were present in the meat.
A major obstacle in the mass utilization of algae as sources of renewable fuel is the high cost of extracting the oil from inside the cells which results in destruction of the cell. To sidestep this obstacle, we would like to use Botryococcus braunii, an algae that lives in colonies and secretes oil into a communal pool which lies outside the cells. Our goal is to develop methods to extract the oil from the colonies with minimal damage to the cells, such that a continuous ‘milking’ process is achieved. The first step is acquiring the Botryococcus braunii and cultivating it continuously in the lab, and then comes the fun part, experimenting with oil extraction techniques. People are welcome to come up with their own ideas and implement them in the lab. Get your thinking caps on and we’ll see you in the lab! If you or someone you know can donate some Botryococcus braunii, please contact us! For now we are fundraising for $250 to purchase a strain.
At the mini maker last year we prepared some differently colored E. coli with Synbiota and their Rainbow kit and gave members of the public a chance to design their own art pieces with it!