That we detected this effect of the wrky17 mutation using a transient T-DNA expression assay indicates that increased VIP1 expression affects the early steps of the infection process, i.e., those that occur prior to T-DNA integration and stable expression.
Although both platforms demonstrated high specificity in DNA binding and gene expression assay, there were several differences.
Results obtained using large scale gene expression assay by DNA microarrays are currently validated using more time consuming techniques such as real-time RT-PCR, Northern blotting, in-situ hybridization and ribonuclease protection assay [ 15].
Before we conducted the expression assay, the genomic DNA of NK 198 was subjected to PCR amplification, targeting a region encompassing the polymorphic sites with primers D-Fw and D-RV to obtain a control template.
We also used DNA expression constructs containing gfp and luciferase for more sensitive assays, but failed to detect any marker gene expression in intact electroporated embryos.
By using the insulin promoter in a luciferase expression assay, they further proposed that a high degree of DNA methylation is associated with low gene expression.
A fragment of the murine ribosomal protein L8 gene (Rpl8) was amplified using the Taqman gene expression assay (Assay ID: Mm00657299_g1) (Applied Biosystems) and used to normalize for input DNA between samples.
d) GUS transient expression assay.
b Transient expression assay in tobacco.
Cis-regulatory motifs of plant promoters have commonly been delineated by the experimental manipulation of DNA segments and reporter gene expression assays [ 3].
