Rapid isolation and characterization of soybean group IX ERF promoters using two different validation tools
Creators:Hernandez-Garcia, Carlos M.
Advisor:Finer, John J.
Contributors:Bouchard, Robert A.
Nemes, Cheri A.
Jones, Michelle L.
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Series/Report no.:Horticulture and Crop Science. Graduate student poster competition, 2009
Promoters are DNA sequences located upstream of gene coding regions that play enormous role in controlling gene expression. Promoters are also important in transgenic crops as they contain the main elements that directly regulate the introduced transgene. Because of their importance, large efforts in our laboratory have recently focused on identification and characterization of plant promoters. Promoters are usually characterized in stably-transformed plants, which take some time to generate. The main goal of this study was to isolate and characterize a soybean ERF group IX promoter family using two new gene expression validation tools. Ten GmERF (Glycine max Ethylene Response Factor) promoters were identified, cloned and initially characterized using quantitative analysis of transient GFP expression in bombarded lima bean cotyledons. Some of the GmERF promoters gave high levels of transient GFP expression, while profiles of transient GFP expression over varies among GmERF promoters. Generation of stably-transformed soybean hairy roots was also evaluated as an additional validation tool for analysis of the GmERF promoter family. Soybean cotyledon explants, inoculated with Agrobacterium rhizogenes, exhibited numerous GFP-positive hairy roots 14-20 days after inoculation. GFP detection in these roots was greatly facilitated by the lack of chlorophyll, which could otherwise interfere with GFP detection. GFP expression intensity was somewhat variable between different events but consistent within events. In summary, 10 different soybean GmERF promoters were isolated and partially characterized in both transiently expressing and stably-transformed tissues.
OARDC/The Ohio State University
United Soybean Board
United Soybean Board
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