Part:BBa_K2449004

cep94A controlled by LacI promoter

cep94A is a cellobiose phosphorylase, which efficiently phosphorylates the cellobiose at its β-linkage, resulting in the degradation of cellobiose to D-glucose and α-D-glucose-1-phosphate.

Exprestion of cep94A

There were conducted an assay to see the expression of the protein coded by Cen94A. It has a weight on 92,7 kDa, it is easely seen on the SDS-PAGE as seen on figure 1 This is on the high copi plasmid;pSB1C3.

Figure 1:A SDS-PAGE of BBa_2449004/cep94A compared with a negative control .The ladder used were SeeBlue™ Plus2 Pre-stained Protein Standard

It was further testet, if the protein could be expressed on a medium copi plasmid;pSB3K3 as seen in figure 2. It would be necessary to have on another plasmid with an other origin of replication, if two plasmids have to transformed into the same bakteria.

Figure 2: BBa_2449004/cep94A on medium copy plasmid pSB3K3 and on the high copy plasmid pSB1C3. The ladder used were SeeBlue™ Plus2 Pre-stained Protein Standard

This SDS-PAGE shows that it is possible to express the protein on pSB3K3.

Groth on cellobiose

The gene cep94A coding for cellobiose phosphorylase could make 'E.coli' live on cellubiose on pSB1C3, this is shown in figure 3. The experiment shows it is not possible to use pSB3K3 plasmid. It is also tested agains No part name specified with partinfo tag.