Team:Hong Kong-CUHK/Project results

After months of wet lab work, we can conclude that our bio-encryption prototype works.

This rci-system is feasible in DH5-alpha strain of E.coli, as supported by extracted plasmid DNA size. It is found that the size of the DNA extracted is consistent with that of DNA stored in the plasmid before extraction. There is no loss of DNA, implying that no large deletion has occurred during the experimental procedure.

In the first trial, we encoded a short message in a single vector, together with two inverted repeats. We designed primers which targets the encoded message either in normal orientation or reverse-complementary orientation. Both sets of primers could be used to generate PCR products, indicating that encoded message exists in both recombinated and normal forms. Sequencing results confirmed the correctness of the PCR product.

Primer sequence to recognize DNA sequence without recombination:

Forward: gCCCAgATCTAgAAAgATggCAATAC

Reverse: TCTCTTATTCTCACTCCACgTACCg

Primer sequence to recognize DNA sequence with recombination:

Forward: CAgATCTAgAAAgATCCACgTACC

Reverse: CTCTTATTCTCACTggCAATACTTTCg

Apart from the above stated, no deletion has been found in the DNA sequence. According to literatures, the recombinase would cause deletion of the sequence upon point mutation. After doing restriction digestion and gel electrophoresis, it is shown that the sequence has a correct size and no deletion is observed.

Yet, there are two mutations noted. However, since high through-put sequencing will be employed, inaccurate bases can be corrected by using majority vote. In short, it can be said that the recombinase can function normally.

To conclude, the whole system of our project is found to be successful and that proves the viability of our project design.