Scaffold attachment regions increase reporter gene expression in stably transformed plant cells.
The yeast ARS-1 element contains a scaffold attachment region (SAR) that we have previously shown can bind to plant nuclear scaffolds in vitro. To test effects on expression, constructs in which a chimeric beta-glucuronidase (GUS) gene was flanked by this element were delivered into tobacco suspension cells by microprojectile bombardment. In stably transformed cell lines, GUS activity averaged 12-fold higher (24-fold on a gene copy basis) for a construct containing two flanking SARs than for a control construct lacking SARs. Expression levels were not proportional to gene copy number, as would have been predicted if the element simply reduced position effect variation. Instead, the element appeared to reduce an inhibitory effect on expression in certain transformants containing multiple gene copies. The effect on expression appears to require chromosomal integration, because SAR constructs were only twofold more active than the controls in transient assays.
Allen
G C
GC
Department of Botany, North Carolina State University, Raleigh 27695.
Hall
G E
GE
Childs
L C
LC
Weissinger
A K
AK
Spiker
S
S
Thompson
W F
WF
D001483:Base Sequence; D002460:Cell Line; D002467:Cell Nucleus / Q000378:metabolism* / Q000648:ultrastructure; D004247:DNA / Q000235:genetics* / Q000378:metabolism / Q000648:ultrastructure; D004742:Enhancer Elements, Genetic; D005784:Gene Amplification; D005786:Gene Expression Regulation; D005966:Glucuronidase / Q000235:genetics; D008957:Models, Genetic; D008969:Molecular Sequence Data; D010947:Plants, Toxic; D012093:Replicon; D012441:Saccharomyces cerevisiae / Q000235:genetics; D014026:Nicotiana; D014170:Transformation, Genetic
D004247:DNA; D005966:Glucuronidase