Team:UChicago/Results
Results
Our team has already made significant strides towards creating the proposed centromeric plasmid vector. We have transformed psB1C3mut into competentE. coli and grown the cultures in LB + CAM plates and liquid media. We similarly transformed psD000 into E. coli as well, along with Pichia pastoris (which we grew up in drop-out arginine plates). Additionally, we have amplified five potential centromeric regions in total from the chromosomal DNA of Pichia pastoris. Our team has already made significant strides towards creating the proposed centromeric plasmid vector. We have grown psB1C3mut in LB+CAM (chloramphenicol), psD000 in LB+CAM and drop-out arginine plates and psD001 in LB+CAM. Additionally, we have amplified five potential centromeric regions in total from the chromosomal DNA of Pichia pastoris. We PCRd yEmRFP from a plasmid in the Glick lab (pOW1-yEmRFP) and ligated it into psD000. In the future, we must sequence psD001 and attempt to grow it in yeast on -Arg plates. Although GeneHackers has made significant progress in our experiment, we still have some parts of our experiment that have not been achieved yet. We need to Gibson assembly various chromosomal sequences into psD001and perform assays of this plasmid in Pichia pastoris. Furthermore, if we are successful in creating the centromeric plasmid, we hope to incorporate the plasmid within the bioengineering and the industrial fields. Here, we want to test the efficiency and practicality of our plasmid within human environments. After these rounds of experimentation, we want to come back to the laboratory and improve our research with this plasmid. Hopefully, we will be able to work toward a final centromeric plasmid that will improve the course of scientific research.