Team:NTU SINGAPORE/Collaborations

Macquaire iGEM Team

Macquaire team had constructed a hydrogenase gene capable of converting glucose to hydrogen gas. The plasmid (BBa_KXXXXXX) that consist of hydrogenase enzyme (Hyd1), ferredoxin, ferredoxin-NADPH-reductase (FNR) and maturation enzyme (HydEF and HydG), in which their expression are co-regulated under pLac promoter.

This year again, our team helped them to evaluate each of the gene expression level by using RT-PCR.

The results of RT-PCR is shown as below:

Formula: 2^-(ΔΔCt) is used to calculate the fold change of gene expression after induction, in which ΔCt(control)1= Ct(uninduced operon genes)- Ct(Chloramphenicol gene)ΔCt(test)2 = Ct(Induced operon genes)-Ct(Induced Chloramphenicol gene)

Finally, use ΔCt(test)2- ΔCt(control)1 to get ΔΔCt.

As shown from the above bar charts, 20mM of IPTG induction has greater fold change of gene expression than 200mM of IPTG induction. It is worth to notice that the RT-PCR efficiency might be different for different genes due to their size of PCR products and the formula used above is assuming the PCR efficiency to be 100% that eventually resulted in 2 copies of products.