Analysis of organ-specific gene expression in soybean
Sundararaman, Vijaya Priya
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Permalink
https://hdl.handle.net/2142/23410
Description
Title
Analysis of organ-specific gene expression in soybean
Author(s)
Sundararaman, Vijaya Priya
Issue Date
1996
Doctoral Committee Chair(s)
Vodkin, Lila O.
Department of Study
Biology
Discipline
Biology
Degree Granting Institution
University of Illinois at Urbana-Champaign
Degree Name
Ph.D.
Degree Level
Dissertation
Keyword(s)
Biology, Molecular
Biology, Genetics
Language
eng
Abstract
Organ specificity is an important criterion to be considered for the expression of foreign genes in plants. For crop improvement by genetic engineering it is critical to introduce genes in specific organs that can be expressed at an appropriate developmental stage. This study involved the analysis of organ-specific control conferred by 5$\sp\prime$ and 3$\sp\prime$ regulatory sequences of the seed-lectin gene of soybean (Glycine max) on the expression of a foreign gene. The reporter gene $\beta$-glucuronidase (GUS), was linked to the 5$\sp\prime$ and 3$\sp\prime$ sequences of the lectin gene and transferred into Arabidopsis thaliana and Nicotiana tabacum. GUS activity assayed on transformed tissues revealed seed-specific expression of GUS under control of the lectin promoter. This expression pattern was found to be conserved among transformed Arabidopsis, and tobacco. These results confirmed that the soybean seed lectin has a strong seed-specific promoter which could be used to successfully drive seed-specific expression of foreign genes.
Another part of this research involved identification of genes that are preferentially expressed during pod development in soybean. To accomplish this, a cDNA library was constructed using mRNA isolated from young pods of soybean. Positive clones identified by PCR were isolated and used to probe RNA blots containing total RNA from a good representation of the different organ types in soybean. Three cDNA clones displayed differential hybridization that were potentially useful for eventually using their promoters to introduce genes that provide protection to the pods against pests and pathogens. The corresponding cDNA inserts VS-16, and VS-51, and VS-53 were sequenced and analyzed.
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