Team:NTU SINGAPORE/Collaborations



Collaboration

Collaborating with these teams allowed us to achieve our goals. It was our pleasure to help them to the best of our ability.

Macquarie iGEM Team


Macquaire team had constructed a hydrogenase gene capable of converting glucose to hydrogen gas. The plasmid BBa_K2300001 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.

IPTG and 20mM of glucose was used to induce the gene expression. Comparison of 1mM IPTG and 20mM IPTG was made and it was found that 1mM IPTG has higher gene expression level. 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.

NUS iGEM Team





















We collaborated with NUS