As EngineerGirl puts it, “When we say engineering principles, we mean the ideas, rules, or concepts that need to be kept in mind when solving an engineering problem.” We knew that engineering principles are very different from scientific research ideas, and the solution to engineering problems pays more attention to specific design and practice. Our team's engineering follows the principle of design-build-test-learnand is committed to the combination of dry and wet labs to improve the efficiency of each cycle.
During the design phase, we fully follow the principle of top-down and carefully consider the recommendations of stakeholders in human practice. From determining abstract biological circuits to selecting actual DNA sequences, from the expected results of the design to the actual tests, we will refine all the relevant details as much as possible at this stage. At the same time, we will work to promote the integration of wet and dry laboratories, by using computers to help wet laboratories model and predict, and designing hardware to optimize wet laboratory workflows and efficiency.
During the construction phase, we will consider the experimental method, the experimental materials, and the expected results in advance, and pay attention to the correlation between the integrity and individuality of the module.
During the testing phase, our focus is on the standardization of the measured data, which we will convert using internationally harmonized units to ensure accurate delivery of information. At the same time, data processing and analysis in the wet lab will also be carried out after communication with the team members of the dry lab.
During the learning phase, we will reflect on the problems that arise and the conclusions that can be drawn from the design-build-test process, and integrate what we think into the next design cycle. We understand that the process of engineering does not happen overnight, so we take an objective look at the implications of each failure or success and use it as much as possible to optimize the engineering of our projects.
Our goal for engineering the sucrose inducible promoter was to characterize the native sucrose inducible promoter and improved it to implement a system that was of practical use when coupled with mazE/F system.
1. The inducible activity should have a wide range to produce a sufficient amount of MazE/F at the beginning of the culture.
2. The promoter should have a lower leaky expression to enable the degradation of the antitoxin mazE.
The first step was to find an inducible promoter and characterized it to base future iteration on.
Next, we needed to find out why the promoter didn’t exhibit the inducible activity. The first concern was the distance between the promoter and the initiation codon (ATG) of the mRFP gene.
In order to use the promoter in our project, we needed to make structural changes to the promoter.
Our goal for part2 was to transform the carbonic anhydrase and ACCBP gene into Bacillus subtilis to obtain the ability to precipitate calcium carbonate with fixed structure outside the cell.
Multi-channel fluorescence detection system had practical significance in many fields. Our goal was to design a multi-channel multifunctional fluorescence detection device to help wet lab to complete the detection of experiments related to luminescent substances.
1.Detection of fluorescent protein concentration
2.Characterization of the oxidation and regeneration process of D-Luciferin
First we needed to complete the preliminary setup of the device to test the oxidation of D-Luciferin.
After determining the shooting parameters, we further detected the reaction of luciferase and D-Luciferin at different temperature conditions, which will helped us determine the thermal stability of luciferase.
Oxyluciferin, which was the product of the luciferase reaction, had a strong inhibitory effect on the firefly luciferase in a manner competitive with firefly luciferin. In the presence of luciferase regenerating enzyme and D-cysteine, oxyluciferin was supplied as the substrate luciferin for next light emission, so that the fluorescence luminescence time of the system could be prolonged.
After characterization of the oxidation and regeneration process of D-Luciferin, we hoped the device to meet the needs of detecting fluorescent protein concentrations.
Linearity was the basis of fluorescent protein concentration detection. We expected to find ranges where there was a strong linear relationship between gray values and fluorescent protein concentrations.
