INVESTIGADORES
RODRIGUEZ TALOU Julian
congresos y reuniones científicas
Título:
Construction of a genetic cassette to express immunogenic dengue virus protein ina plantsystem.
Autor/es:
MARTÍNEZ CA,; GIULIETTI AM; RODRÍGUEZ TALOU J.
Lugar:
Mar del Plata, Argentina
Reunión:
Congreso; XLIII Congreso de SAIB, Noviembre 2007, .; 2007
Institución organizadora:
SAIB
Resumen:
Production of recombinant proteins in plant systems has received
great attention because of the advantages compared with others
production systems. Plant cells can glycosilate and do the posttranscriptional
arrangements need for complex glycoproteins.
Dengue virus genome encodes three structural (C, prM, E) and
seven non-structural proteins. E is an immunogenic glycoprotein
and needs to be directed to the secretory pathway through the Nterminus
signal peptide (SP), to be N-glycosilated. The presence of
an ER retention signal in the C-terminus (KDEL) of some proteins
increases their stability and yield in plant systems. The aim of this
work is to obtain a genetic construct carrying E and CprME protein
with a SP and KDEL cloned in a binary plasmid. E protein was
amplified from plasmid pMT/V5-HisA given by Dr. Gamarnik and
PS was obtained from pCAMBIA1305.2 plasmid by PCR. CprME
was obtained from pMT vector by enzyme restriction. Promoter
(35S) and terminator (T) were obtained by PCR from pMOG843
plasmid. The intermediate constructs were cloned into pGEM T
Vector and sequenced.We obtained the genetic constructions: 35SPS-
E-KDEL-T, 35S-PS-CprME-KDEL-T, 35S-PS-CprME-T and
35S-GRP-E-Term. We cloned these constructions into a
pCAMBIA1305.2 binary vector and we are currently doing the
Agrobacterium-mediated plant transformation.