Difference between revisions of "Part:BBa K3874004"

Latest revision as of 02:35, 21 October 2021

lactoferrin with EGFP

pEGFP-N3- lactoferrin: This gene encoding lactoferrin is located next to a CMV promoter, and its C terminal is connected with an EGFP-tag. We changed the stop codon of lactoferrin and add an EGFP to the C terminal, so this is a new basic part.

Detail information: This year, we are interested in Parts BBa_K3693000 designed by Liu Ziyu in last year’s 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.

Figure 1. The structure of EGFP-N3-plasmid.

Figure 2. The Schematic cartoon of the DNA construct of BBa_K3874004.

Figure 3. The immunofluorescence data of BBa_K3874004.

Figure 4. The western blot data of BBa_K3874004.

Sequence and Features