In the Lab
Our team members selected three possible benzene, toluene, ethylbenzene, and xylene (BTEX) metabolising organisms to focus on - Acinetobacter baylyi ADP1, Rhodococcus sp. 33 and Rhodococcus sp. . Acinetobacter baylyi ADP1 and Rhodococcus sp. have been shown to contain a conventional benzoate degrading pathway.
Cloning of the benABCD cluster and the BenE transporter
Ella was the leader of the cluster portion of the project. She worked to individually clone each gene from the two known clusters and to attach a His-tag. This included the amplification of the cluster and each of the genes from the gDNA and the editing of a vector for the cloning itself. Claudia was in charge of the benE transporter. She also headlined the construction and editing of the vector pTTQRGS6xHisBsaI. She was able to express benE in two strains of E. coli.
Characterising novel BTEX degradation pathways
Rhodococcus sp. 33 was able to utilise benzene as a carbon source through a novel mechanism. Luke investigated benzene degradation pathways in Rhodococcus using various proteomic techniques. This was also used to observe broad cellular responses to the presence of monoaromatic hydrocarbons.
Completing the BenCat sensor
Angus further developed a biosensor that could be used to detect BTEX contamination and degradation in the environment in real time via the expression of fluorescent proteins. The biosensor was initially proposed by the 2020 DeNovocastrians. Previously, the benzoate section of the biosensor was added by Awei Bainivalu. Angus was able to complete the catechol sensing portion of the biosensor and demonstrate the overall activity of the biosensor in E. coli.
Lab maintenance and safety
All of our team members worked collaboratively on the upkeep and maintenacne of the labs we worked in. Additional support with wet work was provided by Ava and Naomi. Safety was an aspect of the project that we all took very seriously. We were all involved in the drafting and submission of the safety form and we maintained clear and open communication about safety expectations throughout the project. The labs we worked within are run by our supervisors Brett Neilan and Karl Hassan and we would like to thank them for their ongoing support.
Outreach and Community
Social media outreach
Naomi served as our social media overlord. She was in charge of running each of our socials from Medium to Twitter to Instagram. Throughout our project she documented our efforts and engaged with the public. We also worked with our University student association in order to develop a synthetic biolgy club. This allowed us to reach out to other students on campus who may be interested in joining the wonderful world of synthetic biolgy and genetic engineering.
Ella and Angus worked to create a podcast to introduce the public to synthetic biolgy. The first episode involved our industry sponsor Samsara and their mission to end plastic pollution with infinite recycling. Luke was in charge of the filming and production of our promotional video, which was then shared on our socials and uploaded to YouTube to spread awareness of our project.
Fundraising
All members of our team with the help of our advisors worked on fundraising. The organisation of fundraising BBQ was lead by Claudia and Niki, with all of our team members coming along to help run the event on the day. This event was within our local area and gave us the added oppurtunity of spreading awareness of both BTEX contamination in local environment and how our team are hoping to fix it. Louis and Dan also helped us reach out to partners such as Samsara for sponsorship throughout the competition.
Thank You!
We would first like to thank all of our advisors; Joachim, Evan, Nicola, Daniel, and Louis, and supervisors; Prof. Brett Neilan and Dr. Karl Hassan. Without them we would not have an iGEM team or a lab to work from and their support and advice throughout the project has been crucial. Secondly, we would like to thank Samsara for their advice, insight, and sponsorship throughout the year. We would also like to thank MPBio for their much appreciated sponsorship. Next, we would like to thank the ARC Centre of Excellence in Synthetic Biology for their support. We would also like to thank the UT Austin and USYD iGEM teams for communicating and collaborating with us this year!
