The deletion of the pif gene improves the biosafety of the baculovirus-based technologies.
Our goal was to improve the biosafety of baculovirus-based technologies by deleting the pif (per os infectivity factor) gene from baculovirus expression vectors. Such a deletion would block transmission in nature without disturbing protein production. A pif deletion mutant of Autographa californica multiplecapsid nucleopolyhedrovirus (AcMNPV) was constructed and its infectivity to two host species was tested by oral or intrahemocoelic inoculation. Virus replication after oral inoculation was monitored using PCR. Oral inoculations with a mixture of the wild type and the pif deletion viruses were carried out. The pif deletion blocked oral infection but it did not hamper infectivity in cell culture. The blockage took place early after inoculation and could not be overcome by mixed inoculations with the wild type. The cat gene was inserted under the control of the polyhedrin promoter in the deletion mutant and the wild type CAT yield was measured in Spodoptera frugiperda insect cells (Sf9) infected with either recombinant. The pif deletion did not hamper CAT production. This deletion significantly improved CAT yields early in the infection. Hence, expression vectors lacking pif may produce higher quality protein. The pif deletion is a simple measure that dramatically reduces the chances of virus spread or gene transfer in nature.
Gutiérrez
Serafín
S
Laboratoire de Pathologie Comparée, INRA UMR1234, CNRS FRE2689 - Université de Montpellier II, 30380 Saint-Christol-les-Alès, France.
Mutuel
Doriane
D
Grard
Nadine
N
Cerutti
Martine
M
López-Ferber
Miguel
M
D000818:Animals; D016367:Baculoviridae / Q000235:genetics* / Q000472:pathogenicity*; D002460:Cell Line; D017353:Gene Deletion; D005818:Genetic Engineering / Q000009:adverse effects*; D012449:Safety; D018411:Spodoptera / Q000821:virology*; D014764:Viral Proteins / Q000235:genetics*; D014777:Virus Diseases / Q000517:prevention & control* / Q000821:virology*; D038301:Virus Inactivation
D014764:Viral Proteins