Part:BBa_K2568006
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 220
- 1000INCOMPATIBLE WITH RFC[1000]Illegal BsaI site found at 1041
Illegal BsaI.rc site found at 1369
Illegal SapI.rc site found at 1168
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