The cooperation with Northeastern University has begun since the summer vocation. We helped them complete a receptor and Working as a very kind and helpful team, they asked helping us in verifying one fuctional protein expression.
We helped NEU-China construct an olfactory receptor Rho-OR1A1. The 64th base pair from the initiation codon of OR1A1 contains an illegal enzyme site (EcoRI), which didn't meet up RFC10 standard. So we designed the primer with synonymous codon and successfully mutated the GAA to GAG.(Figure 1.)
Figure 1. standardized Rho-OR1A1.
In our project, we need various functional vectors for dsRNA production，but the gene sequence complementarity will cause secondary structure, which is impossible to synthsize gene for one time. The task of constructing vectors is heavy.
So NEU-China students helped us construct a hpRNA producing vector ecrL, which speeded up our vectors construction progress. (Figure 2.)
After we meet up through vedio conference with BIT-China, We exchanged ideas and confirmed our collaboration relationship.
In the light of helping each other, we assisted them to build a new type of promoter.
At the same time, they helped us verify one functional protein expression.(Figure 1.)
BIT-China project requires mRFP to characterize the size of the sweetness. Different sweeteners have different gradients of sweetness, they need to get mutations of the inducible promoter Pfus and change its strength.
Figure 1. Mutant gene Pfus.
So we helped them build a mutant gene of Pfus to make their project more responsive.
In our project, pectinase and celluse play an important role in downstream circuits and we want to combine functional dsRNA with these two proteins and let them work together, which will make great sense to our subsequent appllication design.
They successfully verified the expression of pectinase.(Figure 2.)
Figure 2. SDS-PAGE of pectinase.
With their help, our project progress has been promoted.
