Difference between revisions of "Part:BBa K2568006"

Revision as of 11:37, 27 September 2018

Virus receptor Nectin 4 with Kozak sequence and FLAG-tag

The way in which viruses enter into cells is mainly mediated by the host receptor. If a cell can simultaneously express receptors like Nectin 4 protein and TfR protein, and adhesion molecules like heparan sulfate, this cell can be infected by most viruses and is expected to solve the cultivation problem of most viruses.

Our chassis cell MDBK, which is a commonly used cell line in the production of bovine vaccines, has most of the essential receptors and adhesion molecules, and only lacks Nectin 4 protein and TfR protein receptors. Nectin 4 protein is one of the receptors of canine distemper virus (CDV), and its lgV structure forms the strongest binding ability of all receptors with the H protein in CDV. As a kind of cattle cell, MDBK can not be infected by canine virus. But if MDBK can express Nectin 4 protein on the cell membrane, it can be infected by CDV. To achieve the goal of culturing mutiple viruses in the MDBK cell strain, we constructed Nectin 4 biobrick which can be expressed to Nectin 4 protein and transfected it into MDBK cells. When Nectin 4 protein was successfully expressed on the membrane, the CDV can bind to it and invade MDBK cells and then multiply.

Sequence and Features