Contribution
Contribution to Future iGEM Projects
Our project will have great contributions to future iGEM teams, which means that the seminar we hosted motivated knowledge exchange and enhancement between current and potential future iGEM teams, such as our innovative idea being the source of inspiration for future iGEM projects.
Seminar
Our team came up with the idea of hosting a seminar for bio-tech experts and other Taiwanese iGEM teams for sharing ideas and simulating the Grand Jamboree. We invited 6 other iGEM teams from other schools for the seminar, and they included teams from National Chung Cheng University, National Chung Hsing University, Chung Shan Medical University, Kang Chiao Taipei campus, National Tsing Hua University, and Washington High school. Beyond that, we invited Dr. Allen Lien, Dr. Edward Chern, and AIT Speaker Sabrina. They are experts in the fields of biology and medical science, thus their knowledge can help us better prepare for the judging session in the Grand Jamboree. With a total of 92 participants including staff and guests, the seminar will surely benefit every iGem team to develop a deeper understanding of synthetic biology. In the seminar, sessions for team presentations enabled teams to share their ideas and knowledge with each other, and lectures given by experts allowed for students to learn more about biotechnology. This seminar encourages students to broaden their knowledge through knowledge fusion with other teams and learn from experts who have more experience and awareness on the topic.
Innovative idea
Based on the academic paper of “Expression, purification, and characterization of recombinant lumbrokinase PI239 in Escherichia coli”, we came up of allowing our genetically modified E.coli BL21 to produce lumbrokinase when fatty acid is detectedm, with the future potential for a probiotic platform in the human body so that patients wouldn’t have to frequently take in lumbrokinase medication, further aiding prevention or treatment of cardiovascular diseases. The innovative aspect of our project lies in the ability of it being a new method of potential treatment for diseases that include insertion of a genetically modified E.coli into the human body, which aims to produce different levels of protein. Since our experiment is successful, this idea will encourage future iGEM participants to refer to our idea of a bold and courageous innovation compared to current methods of treatment, and this idea will encourage future iGEM teams to think outside the box.
Adding new parts to parts registry
Our team has uploaded several new parts to the igem parts registry. One of them is BBa_K4673029, which is the AmpR
AmpR is a transcriptional regulator regulating the expression of the Citrobacter freundii ampC β‐lactamase gene. It belongs to the LysR family of transcriptional regulators that typically exhibits autoregulatory behavior by repressing its own expression (Ryuichi et.al, 2017). AmpR promoter serves as a critical element in the blue-white screening process, functioning as a binding site for RNA polymerase and leading to transcription of ampicillin resistance genes(b-lactamase coding region). Generally speaking, it is difficult to discern whether each white E.coli DH5α colony comprises the plasmid with the target vector after conducting blue-white screening. However, ampicillin promoter and resistance help. In our team’s project, we utilized the ampicillin promoter for the ampicillin resistance on the pGEM-T EASY vector as the second screening mechanism that can be used to confirm whether the surviving colonies in the results of blue-white screening took up the plasmid that contains the pGEM-T EASY vector during the transformation. In addition to the β-galactosidase marker, most cloning plasmids contain genes that confer resistance to an antibiotic such as ampicillin. Adding ampicillin in the culture medium should prevent bacteria that doesn’t contain the modified plasmid from growing, thereby offering evidence that the white colonies in the culture medium contain the plasmid with target vector. That is to say, the colonies with well-transformed vectors are considered to survive in the presence of ampicillin while the ones without could not function under the condition.
The source of the part:
Nakano, R., Nakano, A., Yano, H., Okamoto, R., Ryuichi NakanoDepartment of Microbiology and Infectious Diseases, N. M. U., Akiyo NakanoDepartment of Microbiology and Infectious Diseases, N. M. U., Hisakazu YanoDepartment of Microbiology and Infectious Diseases, N. M. U., & Ryoichi OkamotoDepartment of Microbiology, K. U. S. of M. (2017, August 30). Role of AMPR in the high expression of the plasmid-encoded AMPC β-lactamase CFE-1. mSphere. https://journals.asm.org/doi/10.1128/msphere.00192-17 Design considerations: (105 bp) Ampicillin promoter is located between T7 terminator and ampicillin resistance in the pET-22b LK vector, and located between the b-lactamase coding region and f1-origin in the pGEM-T EASY vector.