The left graph shows the standard RNA of miracle fruits, and the right graph shows the RNA of miracle fruits that we gain. The DNA molecular weight showed in two graphs matches each other, confirming that we actually collect the authentic RNA of miracle fruit.
We designed the PCR primer of miracle fruit’s cDNA according to primer principle and our seqeuncing result.
- F tcccccggg ATGAAGGAAT TAACAATGCT CTC
- R ggggtaccTTAGAAGTATACGGTTTTGTTG
This is the result of RT-PCR. There were 7 tubes of PCR products, but only two tubes are usable. The seqeunce result of this PCR operation is matched to the standard miracle fruit’s seqeunce, which means the RT-PCR operation is successful, and there is no mutation of base pair.
We digested the DNA we gained, then connect it to T-vector. After that, we transfer the re-constructed plasmid into the competent cell of E.coli.
Smear the E. coli with re-constructed plasmid on the plate with Ampicillin, select the E. coli that could grow on the plate normally. Then we do colony PCR of the selected E. coli. After the PCR operation, the DNA of miracle fruits that shows amplification is confirmed to be positive clone.
Connect the competent cell of lactic acid bacteria to green fluorescent protein. After fostering, the green fluorescent is expressing, confirming that competent cell of lactic acid bacteria is usable.
We collected the plasmid of E.coli. After the digestion, we reconstructed the digestive fragment of pve5523 with the plasmid of E.coli.
We transferred the reconstructed plasmid into E.coli and foster the E.coli. After that, we collected the plasmid of that E.coli and transferred the plasmid into the competent cell of lactic acid. Finally, we got the trans-genetic lactic acid bacteria.
The Sds-page examine for lactic acid bacteria
