- We designed and ordered the gBlocks for 5 genes, which encode for proteins involved in the hydrogen production (hyd1, ferredoxin, hydG, hydEF).
- We attempted to incorporate the gBlocks into BioBricks, which were then transformed into E.coli. Once all the genes can be expressed in E.coli, this will facilitate the production of hydrogen gas.
- Analyse and optimise the genes that are needed to be synthesised.
- Digest and ligate gblocks into Biobricks, , digest check and sequence confirm.
- Digest and ligate Biobricks together via 3A assembly.
- Due to the size of hydEF, the gene was synthesized as 2 gBlocks, incorporated into Biobricks, and brought together by 3A assembly.
- We successfully inserted and confirmed 3 Biobricks (hyd1, hydEF and ferredoxin).
Fig 1. A gel consisting of EcorI/PstI and EcoRI/SpeI (only for hydG) double digests from all the Biobricks. The expected bands were hyd1 (1773bp), ferredoxin (1672 bp) and hydEF (3611 bp), and this was observed in the gel. hydG was not sequence confirmed.