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Chx Promoter::Gus Information

Aim: To examine their precise gene expression, each AtCHX gene promoter region immediately upstream of the ATG start codon was transcriptionally fused with GUS to generate the CHX::GUS reporter.

Brief Protocol: Promoter fragments were amplified by PCR from Col-0 genomic DNA isolated from 3-week-old seedlings using the Expand High Fidelity PCR system (Roche, Mannheim, Germany). The primers used are listed below. For excessively long intergenic regions, only the ~2KB region immediately upstream of ATG was amplified. PCR fragments were made by using the restriction enzyme cut sites of the primer regions (highlighted in blue). These fragments were then verified by PCR and cloned/subcloned into various plasmids in order to create the final GUS constructs. For more specific information and the plasmids used for each specific CHX gene, please read our 2004 Plant Physiol paper.

Legend:
R.E. Site = Flanking restriction enzyme site.

Prom. Size = Promoter size (bp) created by using the listed primers. The number listed in parentheses is the actual intergenic region size for comparison.

(N)PF/PR = (New) Primer Forward/Reverse.