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The pLSG-IBA162 vector is designed for high-level expression of target proteins with an N-terminal FLAG-tag and C-terminal Twin-Strep-tag® in insect cells. The gene transfer into the polyhedrin gene locus of AcMNPV DNA is achieved by homologous recombination, and the vector carries a polyhedrin promoter. Co-transfection with BacPAK6 linearized AcMNPV DNA (Clontech) or with circular flashBAC modified AcMNPV DNA (Oxford Expression Technologies) allows the generation of recombinant baculovirus at very high efficiency through reconstitution of an essential gene (ORF 1629) and elimination of wild-type virus to a great extent. For selection and propagation in E. coli, the vector carries an ampicillin resistance and ColE1 origin of replication (pUC).
It should be noted that cloning into pLSG-IBA Acceptor Vectors compulsorily requires the restriction enzyme Esp3I since no other MCS for the integration of a gene of interest is available. In addition to the direct cloning of the gene of interest into pLSG-IBA vectors with Esp3I, another option via a so-called Entry Vector is possible.
