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GrainGenes Reference Report: PMB-49-45

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Reference	PMB-49-45
Title	Matrix attachment regions (MARs) enhance transformation frequencies and reduce variance of transgene expression in barley
Journal	Plant Molecular Biology
Year	2002
Volume	49
Pages	45-58
Author	Petersen K Leah R Knudsen S Cameron-Mills V
Abstract	Summary: Nuclear matrix attachment regions (MARS) are defined as genomic DNA sequences, located at the physical boundaries of chromatin loops. They are suggested to play a role in the cis unfolding and folding of the chromatin fibre associated with the regulation of gene transcription. Inclusion of MARs in transgene cassettes enhances their expression and reduces position-effect variations in the transgenic host. The present study is the first to investigate the influence of MAR sequences on transformation frequencies and transgene expression in barley, which is highly relevant to the future improvement of this crop by biotechnology. Two plant MAR sequences were tested both for their ability to bind to the nuclear matrix of barley leaf nuclei and to regulate the expression of a reporter gene in transgenic barley. Competitive in vitro MAR binding assays with the 520 bp P1-MAR from soybean and the 516 by TBS-MAR from petunia revealed that only the P1-MAR had specific binding affinity for barley nuclear matrices. The barley transformation frequency with the uidA reporter gene was increased 2-fold when the gene was flanked with either the P1-MAR or TBS-MAR, while the gene copy number was strongly reduced. The presence of P1-MAR sequences increased the mean activity and reduced the variance in expression of a co-integrated reporter gene in barley consistent with the proposed model of MAR activity
External Databases	Pubmed: 12008898
Keyword	[ Hide all but 1 of 55 ] abilities assay attachment attachment region barley barley leaves barley transformation beta-glucuronidase biotechnology booster sequence boundaries chromatin copies copy number dna dna fragment fibre frequencies gene gene transcription gene-fusion marker gene-transcription genomic dna host improvement in vitro mar mars matrix matrix attachment region model nuclear matrix nuclei petunia petunia hybrida promoter regulation reporter reporter gene scaffold sequence simple integration patterns soybean tobacco plants transcription transformation transformation frequency transgene transgene expression transgenic transgenic barley uida unfolding variance vitro

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