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The influence of matrix attachment regions on transgene expression in Arabidopsis thaliana wild type and gene silencing mutants
Authors:Miguel F. C. De Bolle  Katleen M. J. Butaye  Inge J. W. M. Goderis  Piet F. J. Wouters  Anni Jacobs  Stijn L. Delauré  Ann Depicker  Bruno P. A. Cammue
Affiliation:(1) Centre of Microbial and Plant Genetics, Katholieke Universiteit Leuven, Kasteelpark 20, B-3001 Leuven, Belgium;(2) Department of Plant Systems Biology, VIB / Universiteit Gent, Technologiepark 927, B-9052 Gent, Belgium
Abstract:Many studies in both animal and plant systems have shown that matrix attachment regions (MARs) can increase the expression of flanking transgenes. However, our previous studies revealed no effect of the chicken lysozyme MARs (chiMARs) on transgene expression in the first generation transgenic Arabidopsis thaliana plants transformed with a β-glucuronidase gene (uidA) unless gene silencing mutants were used as genetic background for transformation. In the present study, we investigated why chiMARs do not influence transgene expression in transgenic wild-type Arabidopsis plants. We first studied the effect of chiMARs on transgene expression in the progeny of primary transformants harboring chiMAR-flanked T-DNAs. Our data indicate that chiMARs do not affect transgene expression in consecutive generations of wild-type A. thaliana plants. Next, we examined whether these observed results in A. thaliana transformants are influenced by the applied transformation method. The results from in vitro transformed A. thaliana plants are in accordance with those from in planta transformed A. thaliana plants and again reveal no influence of chiMARs on transgene expression in A. thaliana wild-type transformants. The effect of chiMARs on transgene expression is also examined in in vitro transformed Nicotiana tabacum plants, but as for A. thaliana, the transgene expression in tobacco transformants is not altered by the presence of chiMARs. Taken together, our results show that the applied method or the plant species used for transformation does not influence whether and how chiMARs have an effect on transgene expression. Finally, we studied the effect of MARs (tabMARs) of plant origin (tobacco) on the transgene expression in A. thaliana wild-type plants and suppressed gene silencing (sgs2) mutants. Our results clearly show that similar to chiMARs, the tobacco-derived MARs do not enhance transgene expression in a wild-type background but can be used to enhance transgene expression in a mutant impaired in gene silencing. Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users. Miguel F.C. De Bolle, Katleen M.J. Butaye Contributed equally to this work
Keywords:Matrix attachment regions  Transgene expression   Arabidopsis thaliana    Nicotiana tabacum   Gene silencing mutants  High-throughput screening
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