Improvement to the DspB gene used by Team CMUQ of 2017
This year, we are interested in Parts BBa_K3693000 designed by Liu Ziyu in last year鈥檚 iGEM competition. This is a part encoding lactoferrin, which is a major iron-binding protein in breast milk and body secretions with an antimicrobial activity, making it an important component of the non-specific immune system. This protein is essential for human children in their development process. However, there are some mother who are difficult in produce breast milk, their babies are not easy to get this protein. So, we wonder whether we can modify our cells to produce this protein in a large scale to ensure the supply
As a first improvement step, we need to test whether this protein is compatible with the mammal expression system. So, we put it in a cell expression vector pEGFP-N3 to construct pEGFP-N3- lactoferrin.
This gene encoding this protein is located next to a CMV promoter, and its C terminal is connected with an EGFP-tag. After we transfected into HeLa cells, we see obvious fluorescence to confirm this protein can be expressed in human cells. So, here we improved this part to a mammal expression plasmid and add an EGFP-tag onto it. We also confirmed this protein is compatible to express in our cell factories.
