Part:BBa_K5142031:Design
Vaccinia virus A27L
- 10INCOMPATIBLE WITH RFC[10]Illegal prefix found in sequence at 1
Illegal suffix found in sequence at 354 - 12INCOMPATIBLE WITH RFC[12]Illegal EcoRI site found at 1
Illegal SpeI site found at 355
Illegal PstI site found at 369
Illegal NotI site found at 7
Illegal NotI site found at 362 - 21INCOMPATIBLE WITH RFC[21]Illegal EcoRI site found at 1
- 23INCOMPATIBLE WITH RFC[23]Illegal prefix found in sequence at 1
Illegal suffix found in sequence at 355 - 25INCOMPATIBLE WITH RFC[25]Illegal prefix found in sequence at 1
Illegal XbaI site found at 16
Illegal SpeI site found at 355
Illegal PstI site found at 369 - 1000COMPATIBLE WITH RFC[1000]
Design Notes
The A27L protein is a crucial viral protein that plays essential roles in virus attachment, membrane fusion, intracellular transport of the virus within the host cell, and the formation of enveloped virion particles. The protein encoded by the A27L gene is necessary for the cell-to-cell transmission of viral particles. Deleting the A27L gene significantly impairs the virus's ability to spread between cells, resulting in substantially reduced plaque size. Therefore, we chose the A27L protein as the target for our experiments. Without an artificial translation system, the modified virus cannot synthesize complete A27L, leading to a drastic reduction in EEV formation, which greatly decreases the infectiousness and pathogenicity of the modified virus. Viruses unable to synthesize complete A27L mostly remain at the IMV stage, unable to spread over long distances, thereby ensuring the safety of the experiment and its applications. Only when an artificial translation system and unnatural amino acids are added can the modified virus synthesize complete A27L.
Source
The cds of A27L was cloned by PCR amplification from the genome of vaccinia virus (GenBank ID: JX489136.1).