Team:HK SSC/Results
We have successfully constructed our 12kbp plasmid and expressed the dCas9 protein. Here are the results:
1. Plasmid construction
After various restriction enzyme cuts and ligation, we have successfully constructed our 12kbp plasmid.
2. Successful Transformation
We have successfully transformed our plasmids into Microcystis and spread onto selective plates, showing that the plasmid we constructed is compatible in both E.coli and Microcystis. Besides, our transformed cells demonstrated kanamycin resistance. These are living colonies as we checked the plates under microscope every day. We ensured that the cells were not only dead cell bodies buy transfering them to new selective plates once in two weeks.
Fig2. Microcystis Aeruginosa UTEX2388 transformed with our 12kbp plasmid spread onto a Kanamycin plate
Fig3. E.coli DH5α transfromed with our 12kbp plasmid and spread onto a Kanamycin plate.
3. Plasmid expression
We used Microcystin-LR detection kit (WRZ001) from 天河綠洲. We believed that our dCas9-sgRNA complex has been successfully been expressed. This is because the Microcystin-LR concentration in our transformed Microcystis was lower than 0.002mg/L, while our control set ups have a higher concentration.
