Part:BBa_K2568007

Virus receptor TfR with Kozak sequence and His-tag

The way how viruses enter into cells is mainly mediated by host receptors. We have established an association between virus Baltimore subtyping and host receptors, and found 4 internalization receptors and 2 attachment molecules that are necessary to mediate the entry of Baltimore types I to V. Our chassis cell MDBK has most internalization receptors and adhesion molecules, but lacks Nectin 4 and TfR to enable the production of most of the commonly seen animal viruses.

TfR is one of the receptors that mediates the entry of canine parvovirus (CPV). As a kind of cattle cell, MDBK cannot be naturally infected by canine viruses, but can become sensitive to CPV infection if TfR was functionally expressed on cell surface. Therefore, we are motivated to construct the TfR biobrick to achieve the goal of cultivating multiple viruses in MDBK cells..

Characterization

Nucleic acid gel electrophoresis

After constructing the plasmid with TfR, we conducted the nucleic acid gel electrophoresis and DNA sequencing to ensure the accuracy.

Figure 1. Nucleic Acid Gel Electrophoresis M: DL10000 DNA Marker; 1: PLVX-TfR. The 8787 bp band: PLVX plasmid backbone; the 1570 bp band: TfR fragment.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  COMPATIBLE WITH RFC[23]
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal AgeI site found at 721
  
• 1000
  COMPATIBLE WITH RFC[1000]