ISHS


Acta
Horticulturae
Home


Login
Logout
Status


Help

ISHS Home

ISHS Contact

Consultation
statistics
index


Search
 
ISHS Acta Horticulturae 961: VII International Symposium on In Vitro Culture and Horticultural Breeding

POWER AND LIMITATIONS OF DICISTRONIC VECTOR SYSTEMS FOR PLANT TRANSFORMATIONS

Authors:   L.A.I. Vaas, M. Marheine, E. Heine-Dobbernack, H.M. Schumacher, H. Kiesecker
Keywords:   IRES efficiency, expression monitoring, Nicotiana benthamiana, vector architecture
DOI:   10.17660/ActaHortic.2012.961.67
Abstract:
Dicistronic vector constructs harboring an internal ribosomal entry site (IRES) element facilitate the coordinated co-expression of physically independent proteins, as, e.g., a target protein along with a reporter protein. Dicistronic vector constructs have been studied intensively in mammalian cell systems, but their efficiency and applicability in plant cell systems is much less known. The influence of the 1st cistron nucleotide sequence on the translational expression level of the reporter gene encoded by the 2nd cistron was investigated. The testing set consists of the plant viral TMV IRES element and the mammalian viral polio IRES element in combination with either the Atnhx gene, coding for a Na+/H+ antiporter, or the gus gene. Using the resulting four combinations the impact on a firefly luciferase (luc) as reporter gene was studied at the transcriptional, translational and phenotypical level. In order to discriminate between nucleotide sequence of the 1st cistron and its gene product, we introduced a frame shift in the 1st cistron coding sequence. The sequence of the IRES element strongly influenced the translational reporter gene expression present in the 2nd cistron. The results show that the dicistronic vector construct has to be carefully designed and tested for each individual application to obtain the desired level of translational expression of the reporter gene.

Download Adobe Acrobat Reader (free software to read PDF files)

961_66     961     961_68

URL www.actahort.org      Hosted by KU Leuven      © ISHS