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Dicistronic binary vector system—A versatile tool for gene expression studies in cell cultures and plants

Paper ID Volume ID Publish Year Pages File Format Full-Text
24566 43527 2010 8 PDF Available
Title
Dicistronic binary vector system—A versatile tool for gene expression studies in cell cultures and plants
Abstract

Dicistronic binary vector constructs based on pGreenII vectors for Agrobacterium mediated gene transfer alleviate the translational expression monitoring of a target gene in plants. The functionality of the transformation vectors was proven by marker gene constructs containing a mannopine synthase promoter (p-MAS) fused to a beta-glucuronidase (gus) gene followed by an internal ribosome entry site and a firefly luciferase (luc) gene. The cap-dependent translation of a physically independent target protein can be monitored by the cap-independently co-translated luciferase, because both mRNAs are located on the same strand. Among three different IRES elements, the tobamo IRES element showed highest activity in transient expression. As a proof of principle for physiological studies the gus gene was replaced by a sodium antiporter gene (Atnhx1). Comparative studies with Atnhx1 transgenic luc expressing tobacco cell cultures and pea plants (Pisum sativum L.) showed improved salt tolerance in relation to their wild type counterparts grown under corresponding conditions. A coincidence of the luc gene expression and increased sodium chloride tolerance is demonstrated by measurement of luminescence and cell growth.

Keywords
Agrobacterium tumefaciens; Dicistronic; IRES; Salt tolerance; Cell culture; Pisum sativum
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Dicistronic binary vector system—A versatile tool for gene expression studies in cell cultures and plants
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Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Biotechnology - Volume 145, Issue 1, 1 January 2010, Pages 9–16
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering
Get Full-Text Now
Don't Miss Today's Special Offer
Price was $35.95
You save - $31
Price after discount Only $4.95
100% Money Back Guarantee
Full-text PDF Download
Online Support
Any Questions? feel free to contact us