Collaboration

During our iGEM project, we reached out to many teams that were working in plastic degradation or oil dispersion and found a meaningful collaboration with UIUC Illinois. Through this collaboration, we were able to troubleshoot with each other and learn more about each other’s interesting projects.

The UIUC Illinois iGEM Team is working on increasing the efficiency of PETase. Through computational planning and further testing in the wetlab, the UIUC Illinois team will identify mutations that are beneficial for plastic degradation.

The plastics portion of our project focuses on using the P1 bacteriophage as a vector to deliver the plastic degrading enzymes, PETase and MHETase, to bacteria in order to degrade plastic in the environment.

Given that both of our teams are working with PETase, we saw a great potential to collaborate. This collaboration manifested through the troubleshooting of the His-Tag purification of PETase.

July 20

Initial Contact

While taking a look at what other iGEM teams were working on, UIUC's project caught our eye. We thought we had similar goals, so we reached out to them to see if they were interested in working together.

July 25

Our First Meeting

During our initial meeting with UIUC Illinois, we learned more about their project and discussed ways in which we could collaborate. We decided to keep in contact with each other, given that we shared similar goals of purifying PETase. We also shared the assays we would use to assess plastic degradation by purified protein.

July 26

Troubleshooting

As wetlab work picked up, both of our teams encountered difficulties with the induction of PETase with IPTG. Through email communication, we shared papers regarding the plasmid ordered and discovered that PETase may be toxic to BL21 cells, which we were both using as our host. Thus, both of our teams came to the conclusion that we should switch to c41(DE3) and T7 lysY Iq cells that were mentioned in the paper.

Sept 10

Meeting

we discussed our protocols for inducing PETase with IPTG. Our team also shared some insight we received from a graduate student who worked with purifying PETase. For example, we learned that PETase is not stable beyond 30℃ and therefore specific parts of our induction procedure should be changed.

Sept 20

Induction

UIUC Illinois successfully induced PETase using c41 cells after troubleshooting! The UT Austin team has also received purified enzyme for their assays.

Summary:

Overall, our collaboration with UIUC Illinois was incredibly insightful as we were able to troubleshoot many aspects of His-Tag purification with PETase. By switching to c41(DE3) cells and sharing advice we received from each other, we gained key insights that helped with protein purification.