Attributions

The 2018 WPI iGEM Team would like to extend a sincere thanks to all those who contributed to the completion of our project! The team started working in D term, or late May, of 2018 and continued into October of 2018.

Team Members

What follows is a summary of the contributions of each team member.

Bailey Sweet:

In the lab, Bailey worked to purify GFP, IAFGP, and ZeAFP from BL21 strains of E. Coli. To prove that the purification was effective, she worked to run concentration assays as well as SDS page gels as a way to verify the size of the proteins that were purified. Bailey then ran two different Crystal Violet Biofilm assays using the purified protein to determine if they reduce biofilm formation on well plates. On the iGEM team, Bailey was in charge of running the iGEM social media accounts. Twitter and Facebook were used to promote our work, art, and introduction of the iGEM team to the general public.

Kylie Sumner:

In the lab, Kylie performed E. coli transformations, did Gibson assembly cloning, made competent E. coli of various strains, ran more DNA and protein gels than any person should ever have to, and tested the effect of different antifreeze proteins on the formation of biofilm growth. Kylie was also the Collaborations Chair for the team this year.

Alex Rebello:

In lab, Alex was involved in transforming E. coli strains with different proteins, including the various antifreeze proteins tested in this project. He performed Gibson assembly cloning and of EMG competent cells. Alex ran agarose gel electrophoresis to visualize DNA fragments from the samples. Alex also helped with the testing of the gene gun using lettuce and GFP. Alex served as the Integrative Practices Chair for the project and was responsible for reaching out in the experts of the fields of agriculture, GMO regulation, and transgenic plants.

Camille Pearce:

In the lab, Camille was involved in transforming E. coli strains with different proteins, including the various antifreeze proteins tested in this project. She performed Gibson assembly cloning and made competent EMG, DH5a, NCTC and BL21. She was heavily involved in running biofilm assay plates to test the antifreeze proteins’ abilities at disrupting and preventing biofilm growth. Camille ran agarose gel electrophoresis to visualize DNA fragments from the samples as well as his-tag gels to visualize the expressed proteins from the transformations. Camille also held the position of Content Master for the team’s wiki page.

Mei Hao:

In the lab, Mei participated in the AFP-coated lettuce experiments and the mastermind behind the design and execution of the gene gun. She also performed PCR reactions, DNA extractions, and helped maintained our plate stocks and reagents for experiments. As the public relations and graphic design chair, she also aided in the design of the wiki and public presentations documenting experiments.

Beck Arruda:

In the lab, Beck Mei participated in the AFP-coated lettuce experiments and aided in the design and testing of the gene gun. He also participated in developing assays for measuring biofilm and helped maintained lab stocks. As the Web Master, he was the designated guru for all things HTML and helped designed all branding and logos for the team.

Arth Sharma:

In the lab, Arth tested the antimicrobial properties of curcumin (the active ingredient of turmeric) in biofilm assays. With subcloning and protein extraction experiments, he aided in preparing cultures, and measuring sample concentrations. Serving as Outreach and Engagement Chair, he aided the organization and documenting of public demonstrations and workshops in our lab.

Faculty

A special thanks to the WPI faculty who have facilitated, assisted, and encouraged the ongoing research of iGEM and helped make the project possible.

Dr. Natalie Farny:

Advisor of WPI iGEM. Assisted in acquiring of plasmid vectors, reagents, ascertaining protocols, and in subcloning genes.

Mr. Mihail Bocka:

WPI Lab manager. Provided supplies and reagents for our experiments when needed.

Dr. Lou Roberts:

WPI teaching staff. Assisted in electroporation of NCTC strain.

Dr. Mike Buckholt:

WPI teaching staff. Aided in the usage of lab instruments.

Dr. Jill Rulfs:

WPI teaching staff. Provided assistance in troubleshooting broken equipment and gathering necessary lab supplies.

Project Inspiration

With the recent outbreaks of pathogenic bacteria on leafy crops such as lettuce, we sought to address the problem with the research conducted by the 2015 WPI iGEM team. They had confirmed the antibacterial capacity of antifreeze proteins in disrupting biofilm development. Moreover, with the growing societal embracement of natural compounds to combat common pathologies, we assessed the utility of the flavonoid curcumin as an antibacterial.

Supporting Research

We would like to say thanks to Kayla DeSanty and Mitra Marvasti-Sitterly for protocols and insights in their major qualifying project report, "Using Biofilms for the Detection and Bioremediation of Arsenic,"

Outside Institutions

We would like to also express thanks to scientists outside our lab for contributing their knowledge to our experiments.

Dr. Robert Wick:

Plant pathologist. He provided information on plant metabolism, common pathogenic bacteria on crops, plant epidemiology, and safe farming practices.

Dr. Patricia Stapleton:

Political scientist. She provided insight into current food regulations, public opinion of GMOs, and plant classification.

Dr. Susan Roberts and Michelle McKee:

Chemical Engineers. They provided insight into gene gun usage for plant modification and transgenic plants.

Glenn Stillman:

Biochemist and farmer. He described current farm practices, showed us his farm and explained his business model, and provided context for the adoption of a GMOs and pesticides.

iGEM Teams

We would like to extend a deep thanks to the Pasteur Paris iGEM team for their biofilm assay protocol!

Plasmid Source

The pGFPGUSPlus we used was a gift from Claudia Vickers (Addgene plasmid # 64401).

Financial Support

Without the generous support from the following folks, our project wouldn't be possible: WPI Dean of Engineering, Winston Soboyejo, the WPI center for global public safety, Dept. of Biology and Biotechnology, Dean of Arts and Sciences, Jean King, IDTDNA, and BioBasic.