Measurements
Our project this year aims to make the strains with target genes grow faster than those who don’t. Therefore, quantitative data is a must in order to confirm the difference between experiment strain and the control strain. Moreover, since the final part may consist of many kinds of transcription switches, these regulatory parts themselves need to be tested as well. In order to finish these tasks, we’ve held a huge amount of quantitative experiments this year, most of them were done with the help of a plate reader or a flow cytometer.
The experiments briefly described below were all based on our design of the gene circuits. To see our designs, click: Link to 2.Design
First of all, we want to test the function of our growth-promoting factor (glucose dehydrogenase, GDH). We want to verify if the gene can increase the K value of the population as well as if the gene has the ability to keep the target genes we actually need. We carried out some delicate experiments and effectively dealt with the numbers. For more information, click here: Link to Result:Specific gene circuit module 1
As for the verifying experiments of the regulatory parts, all of the regulatory proteins were connected downstream of a lac-inducible promoter and florescence proteins were used as an “effector” through all the related experiments. We know that there may be various problems when carrying out the tests, thus every move had been carefully designed and strictly obeyed. We also applied some useful methods dealing with our data to get a more reliable outcome.
For more information, click here: emrR part: Link to Result:Specific gene circuit module 2 and 3 lambda CI repressor part: Link to Result: Universal gene circuit module 5 TGATG: Link to Result: Universal gene circuit module 4
Finally, we want to validate the function of the combined vector and further demonstrate our work. We need to figure out a relationship between the expression level and the growth-promoting effect. We also want to compare our engineered strain with those strains containing antibiotic resistance genes. Various experiments were held in this area. Results were scientifically analyzed and possible improving approaches were properly discussed. For more information, click here: Link (Intergrated Plasmid) Link (Further Improvement)