NDLI: High-level expression of the HIV-1 Pr55gag polyprotein in transgenic tobacco chloroplasts

Content Provider	Springer Nature Link
Author	Scotti, Nunzia Alagna, Fiammetta Ferraiolo, Enrico Formisa, Gelsomina Sanni, Lorenza Buonaguro, Luigi De Stradis, Angelo Vitale, Alessandro Monti, Luigi Grillo, Stefania Buonaguro, Franco M. Cardi, Teodoro
Copyright Year	2009
Abstract	Plants have been recognized as a promising production platform for recombinant pharmaceutical proteins. The human immunodeficiency virus Gag (Pr55gag) structural polyprotein precursor is a prime candidate for developing a HIV-1 vaccine, but, so far, has been expressed at very low level in plants. The aim of this study was to investigate factors potentially involved in Pr55gag expression and increase protein yield in plant cells. In transient expression experiments in various subcellular compartments, the native Pr55gag sequence could be expressed only in the chloroplast. Experiments with truncated subunits suggested a negative role of the 5′-end on the expression of the full gene in the cytosol. Stable transgenic plants were produced in tobacco by Agrobacterium-mediated nuclear transformation with protein targeted to plastids, and biolistic-mediated plastid transformation. Compared to the nuclear genome, the integration and expression of the gag transgene in the plastome resulted in significantly higher protein accumulation levels (up to 7–8% TSP, equivalent to 312–363 mg/kg FW). In transplastomic plants, a 25-fold higher protein accumulation was obtained by translationally fusing the Pr55gag polyprotein to the N-terminus of the plastid photosynthetic RbcL protein. In chloroplasts, the Pr55gag polyprotein was processed in a pattern similar to that achieved by the viral protease, the processing being more extended in older leaves of mature plants. The Gag proteins produced in transgenic plastids were able to assemble into particles resembling VLPs produced in baculovirus/insect cells and E. coli systems. These results indicate that plastid transformation is a promising tool for HIV antigen manufacturing in plant cells.
Starting Page	1109
Ending Page	1122
Page Count	14
File Format	PDF
ISSN	00320935
Journal	Planta
Volume Number	229
Issue Number	5
e-ISSN	14322048
Language	English
Publisher	Springer-Verlag
Publisher Date	2009-02-21
Publisher Place	Berlin/Heidelberg
Access Restriction	One Nation One Subscription (ONOS)
Subject Keyword	HIV-1 Pr55gag Plant factories Plastid transformation Protein accumulation Tobacco Ecology Forestry Agriculture Plant Sciences
Content Type	Text
Resource Type	Article
Subject	Genetics Plant Science

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