Team:NYMU-Taipei/Part Collection
Parts Collection
Odorant Biosensors for Detecting the Scent of Lung Diseases
For the first time in iGEM part collection, we are contributing the best part collection for detecting the scent of diseases using odorant biosensors. The BBa_K3268000 part of our collection is designed to highly express mOR103-15, an olfactory receptor protein that binds to heptanal, in E. coli cells. We have also designed another mOR103-15 expressing part, BBa_K3268001, that can be expressed on the outer surface of E. coli cells. Heptanal can be used as a breath screening marker for lung cancer and tuberculosis disease. Volatile organic metabolites can be generated in many human samples through distinct metabolic pathways. Increasing research evidence indicates that patients’ bodies give off unique Volatile Organic Compounds (VOCs) in different stages of human diseases. Therefore, VOCs are considered as potential biomarkers for performing non-invasive and patient-friendly disease screening in the early stages. This year, NYMU-Taipei iGEM team has taken lung cancer and tuberculosis disease as an example and created intracellularly expressed and surface-displayed odorant-binding proteins to detect and monitor the volatile biomarkers from the patients’ bodies. We have immobilized expressed olfactory receptor proteins to paper device prototypes to detect heptanal which is a specific disease VOC. Here, we provide an example to demonstrate the feasibility of using odorant biosensors to perform non-invasive early disease screening.
Part: BBa_K3268000
mOR103-15
This part codes for mouse olfactory receptor protein, which can bind with heptanal.
Our iGEM team had made a composite module that can highly express any inserted protein-coding genes (see BBa_K3268004). We had used this composite module to highly express mOR103-15 (mouse olfactory receptor protein) in E. coli cells.
- The mOR103-15 coding region was PCR amplified from the pI7-ISH plasmid (https://www.addgene.org/15527/) obtained from Addgene using high-fidelity enzyme KOD-Plus.
- Since the mOR103-15 coding sequence contains a BamHI cutting site, we had designed two PCR primers with HindIII and BglII sites respectively used for this amplification.
Part: BBa_K3268001
Lac-inducible generator of Lpp-OmpA(46-159)-mOR103-15 + His6 tag
This part contains a lac promoter and strong ribosome binding site for lac-inducible expression of the fusion protein of Lpp signal peptide, OmpA aa46-159, and mOR103-15 + His6 tag. This expression should display streptavidin on the cell surface of E. coli.
In order to display mOR103-15 on the surface of cell, we found that the Lpp′OmpA(46-159) hybrid protein can serve as an efficient targeting vehicle for localizing a variety soluble proteins onto the E. coli surface.We got the part BBa_J36850 which is combined with a lac promoter and strong ribosome binding site for lac-inducible expression of the fusion protein of Lpp signal peptide, OmpA aa46-159, and streptavidin wild-type + His6 tag from iGEM kit 2019.
