Team:Missouri Rolla/Safety
Safety
All S&T iGEM members must be trained through our online and in-lab Lab Training Program before contributing to the project in lab. Our Lab Training Manual, which contains a variety of safety and procedure information, and our online lessons are currently being revised to better prepare future lab workers. The Lab Training Program culminates in the "mini-project," where trainees are given a small task and must produce a final product using parts from the iGEM DNA Distribution. Our goal is to train safe and productive lab workers by giving them a chance to try each procedure themselves under the supervision of an experienced member.
We used E. coli DH5α, a non-pathogenic strain, for all assembly and as the chassis for testing. We attempted to express the mevalonate pathway to produce (E)-beta-ocimene, a safe terpene with pleasant scent. We also used Streptomyces roseus, another non-pathogenic strain. The genome of S. roseus contains two genes of interest, leupA and leupB, which are potentially involved in the production of leupeptin, a protease inhibitor. Our final safety form may be found here.
While assembling our parts, we used a lac promoter to control expression. However, a different approach would be taken for use outside the lab. Ocimene indiscriminately inhibits microbial growth, so continuous production in a cave environment would have the potential to significantly impact the microbiome of bats' skin and the cave itself. We are investigating ways of sensing the causative agent of White-Nose Syndrome, P. destructans, as a potential future project. For real-world applications, sensing of the fungus would need to be coupled with either a kill switch or regulation of ocimene production. Even better, ocimene could easily be maintained at a constant concentration within a cave using automatic release, while leupeptin could likely be applied to the bats' skin before they begin hibernation. These applications of our microbial production would prevent both the release of genetically-modified bacteria and disturbing bats during their torpor. Before use for bat conservation, it should also be confirmed that ocimene and leupeptin have no adverse effects on bat species.
Lab photos credit: Bob Phelan, SDELC
