Team:XJTLU-CHINA/Demonstrate
The Results
Our team first tested the expression and solubility of Qbeta replicase which is used to replicate the template later. The results demonstrated the increment of expression and solubility of Qbeta replicase under the expression and effect of EF-Tu and EF-Ts, as mentioned by Gunasekaran (2013). Secondly, the activity and effect of Qbeta replicase is tested by representation Kanamycin resistance in E. coil. The negative strand mRNA with Kanamycin resistance became positive mRNA strands under the effect of replicase and our cells expressed Kanamycin resistance that can grow on the Kanamycin medium.
Thirdly, intron-target mechanism is measured by the length increasing of AKB plasmid. After PCR and agarose gel electrophoresis, the length of AKB plasmid increased 2200bp. Finally, In the igem project of previous year, after screen of more than ten types of RNAT, it turned out A1 the best RNAT that showed most expression difference of eGFP when raised the temperature from 30℃ to 37℃. This year we characterized this RNAT using different reporter gene, mRFP followed by the constitutive promoter J23119. However, this time, rather than presenting difference in different temperature, the mRFP barely expressed in this system.
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