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United States Department of Agriculture

Agricultural Research Service

Title: A Normalized Transient Expression System to Study Transgene Expression

Authors
item Ma, Hongmei
item Griesbach, Robert
item Pooler, Margaret

Submitted to: In Vitro Biology Meeting
Publication Type: Abstract Only
Publication Acceptance Date: February 20, 2005
Publication Date: June 7, 2005
Citation: Ma, H., Griesbach, R., Pooler, M.R. 2005. A normalized transient expression system to study transgene expression. InVitro. 41:43-A.

Technical Abstract: Transient expression has a wide range of applications in molecular biology. It is often carried out by direct DNA transfer rather than Agrobacterium. The expression of genes on the plasmid DNA can be detected very soon. One can easily test a large number of putative important transgenes in a relatively short time period before committing to the most promising candidate gene for stable transformation. However, variables inherent in the tissue culture and bombardment processes make it difficult to compare data from independent transformation events. The goal of this work was to develop a normalized transient expression system by introducing two control reporter constructs along with a construct of interest. The reporter constructs contained either ZEAmay;MYC;Lc or ZEAmay;MYB;C1 (Zea mays anthocyanin regulatory genes) under the control of the CaMV 35S promoter. These two genes are capable of activating anthocyanin biosynthesis in white Phalaenopsis amabilis flowers. The resulting anthocyanin production was used to standardize transient expression. The test constructs included five plasmids, each containing a different promoter fused to a bifunctional GFP:GUS reporter gene. Using the petals of white Phalaenopsis amabilis flowers, the promoter activity was evaluated by transient co-expression of three constructs, one test construct and two control constructs.

Last Modified: 8/27/2014
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