Transient reporter gene (GUS) expression in creeping bentgrass (Agrostis palustris) is affected by in vivo nucleolytic activity |
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Authors: | Basu Chhandak Luo Hong Kausch Albert P Chandlee Joel M |
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Affiliation: | (1) Department of Plant Sciences, University of Rhode Island, 9 East Alumni Avenue, Suite 7, Kingston, Rhode Island 02881, USA;(2) HybriGene, Inc., 530 Liberty Lane, West Kingston, Rhode Island 02892, USA;(3) Department of Cell and Molecular Biology, University of Rhode Island, Morrill Science Center, Kingston, Rhode Island 02881, USA |
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Abstract: | Leaf and callus tissues of a creeping bentgrass cultivar (Penn A4) had high nuclease activities that degraded exogenously added plasmid DNA. When callus tissue was incubated for 24 h with heparin, spermidine, aurintricarboxylic acid or polyethylene glycol, only heparin and spermidine were effective as in vitro nuclease inhibitors, protecting exogenously added plasmid DNA from degradation. When -glucuronidase (GUS) reporter gene activity was evaluated in heparin-treated (0.6%), 14-month old callus following microprojectile bombardment, GUS activity increased 1000-fold compared to equivalent aged untreated Penn A4 callus. Similar enhancement from heparin pretreatment (0.6% or 1.2%) was not observed in 6-month old callus. This is likely due to much higher activities of nuclease in the younger callus. |
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Keywords: | /content/q243m9752tn06n67/xxlarge946.gif" alt=" beta" align=" MIDDLE" BORDER=" 0" >-glucuronidase creeping bentgrass microprojectile bombardment nuclease |
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