Part:BBa_K2568007
Virus receptor TfR with Kozak sequence and His-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. TfR protein is one of the receptors of canine parvovirus (CPV), and it can strongly bind to CPV. As a kind of cattle cell, MDBK can not be infected by canine virus. But if MDBK can express TfR protein on the cell membrane, it can be infected by CPV. To achieve the goal of culturing mutiple viruses in the MDBK cell strain, we constructed TfR biobrick which can be expressed to Nectin 4 protein and transfected it into MDBK cells. We hopes that TfR protein can be successfully expressed on the membrane, the CDV can bind to it and invade MDBK cells and then multiply.
Characterization
Nucleic acid gel electrophoresis
After constructing the plasmid with TfR biobricks, we conducted the nucleic acid gel electrophoresis and DNA sequencing to ensure the accuracy.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal AgeI site found at 721
- 1000COMPATIBLE WITH RFC[1000]
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