REDBIO México 2010

Folio: 476
Tipo de presentación: Cartel
Tema: Otro

An efficient method for the expression of the Bovine Viral Diarrhea Virus Glycoprotein E2 in Plant Cells

Nelson G (1), López J (1), La Torre JL (1), Marconi P (1), Alvarez MA * (1).
* Autor para correspondencia (Corresponding author) malvarez@conicet.gov.ar

RESUMEN (ABSTRACT)
"Bovine Viral Diarrhea is a widespread disease that causes serious mucosal lesions and other clinical manifestations in cattle. It generates a negative economic impact in dairy operations that is between $20 and $160 per adult cow per year. The causative agent is the BVDV, which is a member of the genus Pestivirus. The glycoprotein E2 is the major target of the protective immune response against BVDV infection. Commercially available inactivated and modified-live BVDV vaccines have been extensively used. Even though, there is still a controversy regarding their efficiency. Plants are now gaining widespread acceptance as an alternative platform for vaccine production. Among them, plant cell cultures combine the virtues of the whole-plant systems with those of microbial and animal cell culture, including the implementations of GMP throughout the production pipeline, the precise control over growth conditions, and the independence of outside variables. We have produced two constructs carrying the E2 coding region without its transmembrane domain, under the control of the CaMV35S promoter and T35S terminator, carrying the 2s2 secretory signal, the His-tag, and with or without the endoplasmic reticulm retention signal KDEL. We performed Nicotiana tabacum transformation by using Agrobacterium tumefaciens carrying those constructs. We have obtained tE2 productive transgenic tobacco lines with a percentage of positiveness of 38 %. The establishment of tobacco in vitro lines expressing tE2 with and without the endoplasmic reticulum retention signal KDEL will permit us to make comparative studies of the recombinant protein yields according their place of accumulation (culture medium or biomass)."