Team:Hawaii/Attributions
MAHALO NUI LOA
We are extremely grateful to have received project support from a brilliant crew of advisors and funding from University of Hawaii at Manoa and the National Science Foundation. The team started out the year with some members having little to no lab experience outside of the classroom. We would not have made it through without the countless hours of guidance from an extraordinary cast of supporters. Thank you to Dr. Gernot Presting, Dan Laspisa, and Vishal Negi for invaluable counsel in wet lab and bioinformatic processes and Ryan Shontell for providing critical direction, resources, and advice to our undergraduate students throughout the entire iGEM project. Thank you to all of our team members for pushing the project forward through the literature review period from last fall to the intense summer and fall wet lab period this year. We needed all hands on deck with a small team and appreciate all the sacrifices made to achieve our project milestones. Major roles of our team members are described below.
John Banasihan took charge of the virus-like particle (VLP) assembly process. He cloned our N-terminal constructs into DH5a and BL21, purified proteins for subsequent VLP analysis, optimized buffer conditions for our Gag variant VLPs, and designed time assays to observe VLP intermediates. He actively assisted in all aspects of other lab work.
Fernanda Hennig modelled our VLPs to determine packaging parameters, designed the initial Biobrick primers, and created SDS-PAGE and agarose gels. She assisted in various aspects of the cloning and purification process. In addition, she helped in formatting lab images and documents for recording purposes.
Shelby Roberson made competent BL21 and DH5a cells for our use throughout summer and fall and assisted with cloning attempts of the C-terminal purification tagged constructs into DH5a. In addition, she created the poster for our poster session at the Giant Jamboree.
Jon Tello assisted in the cloning attempts of the C-terminal purification tagged constructs, various aspects of induction, protein purification, and restocking of general lab supplies. He was tasked with sequencing many of our constructs over the summer and prepared the Biobricks (drying and mailing) for shipping.
Gina Watanabe designed gag construct variants and Gibson and non-Gibson primers, led the initial Gibson Assembly and N-terminal and C-terminal cloning attempts, optimized the cloning process with the use of our homemade competent cells, cloned constructs for Biobrick submittal, took charge of cloning the Gag-RFP fusion protein, and actively assisted and organized all aspects of lab work. She strategized to push the project forward throughout and coded the Wiki (code + graphics) with prior experience in coding.
Emily Yang managed the collaboration with Team Dalhousie, assisted with testing the Gibson Assembly method and various aspects of the cloning, induction, and protein purification processes. She was actively engaged in wet lab work over the summer, then helped to add Wiki content while in California during the fall semester.
Thank you to the National Science Foundation and UH Manoa Undergraduate Research Opportunities Program (UROP) for generous funding to conduct and present our project. This is the first observation of virus-like particles (VLPs) in centromeric retrotransposon (CR) elements of Zea mays and will provide one step forward towards characterizing VLPs of this system for potential use in gene therapy and drug delivery. Mahalo for the opportunity to learn and explore a remarkable natural system. We hope our data draws further interest in fully characterizing these centromeric retrotransposable elements.