Team:Georgia State/Safety
Safety
Our Project
The ultimate goal of our project was to create an easy, cost-effective, and sensitive contamination detection
device for iGEM teams using Human Chorionic Gonadotropin (HCG). Initially, we found that Factor C, a protein
within Horseshoe crab blood, is activated by lipopolysaccharides (LPS). In the presence of LPS, Factor C undergoes
auto-catalysis which initiates the cascade of protein activation steps leading to coagulation. To replace this
complicated cascade, we proposed to devise a detection system that signals auto-catalysis of Factor C.
We first tested various chains of HCG at different concentrations to see how they vary in sensitivity to the
pregnancy test "Clinical Guard." Then based on our results, we will pair the most effective chain of HCG with Factor
C. Then we will insert the DNA sequence into pSB1C3 for submission, and pGEX for protein expression. From there
we will purify the protein produced by the pGEX. To produce a detection system we will fuse the purified protein
to the membrane strip and insert it into a saline solution. Then to check for contamination, a pregnancy test strip
will be inserted into the solution and will trigger a response based upon the presence of LPS. With regard to the initial goal, the direction of the lab has changed to focus on developing a universal protein
detection system for low resource labs using HCG via production within bacteria plasmid DNA by way fusion with
Glutathione-S-Transferas. The Georgia State University iGEM lab is located on the 4 th floor of Classroom South in room 443; it is a Biosafety
Level 1 (BSL-1) laboratory. The reagents and organisms used are saved in refrigerator or biosafety cabinets,
respectively. Work is usually conducted in a laminar hood or on an open lab bench. Despite there being no known
pathogenic risks in BSL-1 laboratories, the following precautions were still taken: wearing gloves at all times,
removal of gloves and proper handwashing before exiting the lab, no food or drinks within the lab, tightly
regulating access to the lab, and the inactivation of infection material via autoclaving and hyperchloric acid
(bleach). Prior to entering the lab, everyone had to undergo safety training and tests which include the Right-to-Know
training, Right-to-Know Chemical Specific training, and Hazardous Waste Generator training.
Escherichia Coli DH5a competent cells do not pose a risk to human health; they are classified
as a Risk Group 1 and are non-pathogenic. We do not plan to ever use any resistance strains,
so the bacteria presently in use can be easily be contained within the lab. We have no intention
within this project to inject organisms or reagents into animals or vectors that are to leave the
lab.
Safety and Training in the Lab
Safety and Ethical risks
