Details zum Vektor
| BacPAK6 | |
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Hitchman RB, Possee RD, King LA (2009). Baculovirus expression systems for recombinant protein production in insect cells. Recent patents on biotechnology 3(1):46–54. |
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| The BacPAK System uses the baculovirus Autographa californica nuclear polyhedrosis virus (AcMNPV) to produce target proteins in insect cells. The target gene is inserted into a shuttle vector, which is cotransfected into insect host cells with the linearized BacPAK6 Viral DNA. The specially designed BacPAK6 Viral DNA forces recombination between the virus and transfer vector, resulting in high recombination efficiency. Following recombination, a few viral plaques are picked and purified, and the recombinant phenotype is verified. The newly isolated recombinant virus can then be amplified and used to infect insect cell cultures to produce large amounts of the desired protein. BacPAK6 has an essential gene adjacent to the polyhedrin locus that provides selection for recombinant viruses . Sites for Bsu36 I, which does not cut wild-type AcMNPV DNA, were introduced into the genes flanking the polyhedrin expression locus of BacPAK6. Digesting BacPAK6 with Bsu36 I releases two fragments. The first carries part of a downstream gene, ORF1629, that is essential for viral replication. If the second large DNA fragment recircularizes by itself, the resulting viral DNA will lack an essential part of the genome and be unable to produce viable viruses. | |
AcMNPV genome with modifications: |
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