A transformation booster sequence (TBS) from Petunia hybrida functions as an enhancer-blocking insulator in Arabidopsis thaliana
Several matrix-attachment regions (MARs) from animals have been shown to block interactions between an enhancer and promoter when situated between the two. Since a similar function for plant MARs has not been discerned, we tested the Zea mays ADH1 5′ MAR, Nicotiana tabacum Rb7 3′ MAR and a transformation booster sequence (TBS) MAR from Petunia hybrida for their ability to impede enhancer–promoter interactions in Arabidopsis thaliana. Stable transgenic lines containing vectors in which one of the three MAR elements or a 4 kb control sequence were interposed between the cauliflower mosaic virus 35S enhancer and a flower-specific AGAMOUS second intron-derived promoter (AGIP)::β-glucuronidase (GUS) fusion were assayed for GUS expression in vegetative tissues. We demonstrate that the TBS MAR element, but not the ADH1 or Rb7 MARs, is able to block interactions between the 35S enhancer and AGIP without compromising the function of either with elements from which they are not insulated.
KeywordsMAR Transformation booster sequence 35S promoter/enhancer AGAMOUS second intron Arabidopsis thaliana Enhancer-blocking insulator
Matrix attachment region
Transformation booster sequence
AGAMOUS second intron-derived promoter
35S cauliflower mosaic virus promoter/enhancer
Green fluorescent protein
The authors wish to thank Mr. Dennis Bennett (USDA-ARS, Kearneysville, WV) for his technical assistance, and Dr. Ann Callahan (USDA-ARS, Kearneysville, WV) for critical reading of the manuscript. This study was funded by the USDA-ARS Headquarter 2005 and 2007 classes of postdoctoral grants and a USDA CSREES BRAG grant (2006-03701).
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