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THE PLANT CELL, Vol 7, Issue 5 599-609, Copyright © 1995 by American Society of Plant Biologists
The MAR-Mediated Reduction in Position Effect Can Be Uncoupled from Copy Number-Dependent Expression in Transgenic Plants
L. Mlynarova, R. C. Jansen, A. J. Conner, W. J. Stiekema and J. P. Nap
Department of Molecular Biology, Centre for Plant Breeding and Reproduction Research (CPRO-DLO), P.O. Box 16, NL-6700 AA Wageningen, The Netherlands
To study the role of matrix-associated regions (MARs) in establishing
independent chromatin domains in plants, two transgenes were cloned between
chicken lysozyme A elements. These transgenes were the neomycin
phosphotransferase (NPTII) gene under control of the nopaline synthase
(nos) promoter and the [beta]-glucuronidase (GUS) gene controlled by the
double cauliflower mosaic virus (dCaMV) 35S promoter. The A elements are
supposed to establish an artificial chromatin domain upon integration into
the plant DNA, resulting in an independent unit of transcriptional
regulation. Such a domain is thought to be characterized by a correlated
and position-independent, hence copy number-dependent, expression of the
genes within the domain. The presence of MARs resulted in a higher relative
transformation efficiency, demonstrating MAR influence on NPTII gene
expression. However, variation in NPTII gene expression was not
significantly reduced. The selection bias for NPTII gene expression during
transformation could not fully account for the lack of reduction in
variation of NPTII gene expression. Topological interactions between the
promoter and A element may interfere with the A element as a domain
boundary. In contrast, the GUS gene on the same putative chromatin domain
showed a highly significant reduction in variation of gene expression, as
expected from previous results. Surprisingly, no copy number-dependent GUS
gene expression was observed: all plants showed approximately the same GUS
activity. We concluded, therefore, that dCaMV 35S-GUS gene expression in
mature tobacco plants is regulated by some form of dosage compensation.
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