| Abstract Detail
Mechanisms of Gene Regulation Dhillon, Taniya [1], Chiera, Joseph [2], Finer, John [2]. Comparison of Viral Suppressors of Silencing to Stabilize Transgene Expression. Introduction of the green fluorescent protein (gfp) gene in a transient expression system results in peak GFP expression 24 hrs post-introduction, followed by a rapid decline within 72 hrs. This decline in GFP expression may result from DNA degradation or silencing of the highly-expressed gfp gene. We evaluated six different viral “suppressors of silencing” for their abilities to stabilize GFP expression using a new transient expression system. Suppressor constructs were generated to allow introduction of the suppressor either as a translational fusion with the gfp gene, or on a separate plasmid from the gfp gene. Various constructs were introduced into lima bean (Phaseolus lunatus L.) cotyledons from germinating seedlings, using particle bombardment. Post-introduction, GFP expression was tracked over time using an automated image collection and analysis system. The effects of the suppressors were quantified using image analysis of the resultant GFP expression profiles. The silencing suppressors HCPro and p19 stabilized GFP expression when introduced as 3΄ gfp translational fusions, while p21 and γb stabilized GFP expression when co-introduced with the gfp gene on separate plasmids. The last two suppressors, AL2 and replicase, did not stabilize GFP expression in bombarded lima bean cotyledons. Introduction of the 3΄ gfp fusions of p19 and HCPro into soybean (Glycine max (L.) Merrill) resulted in transgenic cultures and plants, displaying the typical variation in GFP expression. Although no novel phenotype was recovered from the introduction of the HCPro fusion gene in transgenic soybean plants, introduction of the p19 fusion resulted in a downward-leaf curling phenotype, suggesting that the fusions made with HCPro and p19 may differentially affect plant development.
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1 - The Ohio State University, Horticulture and Crop Science, 225 Williams Hall, OARDC/OSU, Wooster, Ohio, 44691, USA
2 - The Ohio State University, Horticulture and Crop Science
Keywords:
GFP
Particle bombardment
Viral suppressors of silencing.
Presentation Type: Plant Biology Abstract
Session: P
Location: Exhibit Hall (Northeast, Southwest & Southeast)/Hilton
Date: Sunday, July 8th, 2007
Time: 8:00 AM
Number: P36041
Abstract ID:1847 |
