pUBI10-GUS Reporter Plasmid

pUBI10-GUS is a reporter plasmid developed for transient transfection of plant (e.g., Arabidopsis thaliana) protoplasts. It employs the β-galacturonidase (GUS) reporter gene from Escherichia coli which has no endogenous expression in plants, but confers stable expression of GUS in plant protoplasts after transfection. This plasmid contains the ampicillin marker for bacterial selection.

Expression of the GUS gene in pUBI10-GUS vector is driven by the Ubiquitin 10 (UBI10) gene promoter from Arabidopsis. The constitutive expression of GUS genes driven by the cauliflower mosaic virus 35S (CaMV35S) promoter increases the metabolic burden in plant, causes a huge waste of material and energy, and thus affects the normal growth and development of the plant. By contrast, the UBI10 promoter not only can efficiently drive the expression of downstream genes, but also can avoid the disadvantages of CaMV35S promoter.

There are several enzyme digestion sites in the upstream and the downstream of the UBI10 promoter in pUBI10-GUS plasmid. Therefore, the UBI10 promoter can be replaced with other promoters to investigate the activation of the promoter of interest.

The pUBI10-GUS vector contains only the components required for transient expression in plants to ensure higher transformation efficiency of protoplasts. This vector cannot be used to obtain stable transgenic plants.

pUBI10-GUS employs the nopaline synthase terminator (NOS-T) from Agrobacterium rhizobium to terminate the transcription by RNA polymerase and regulate the expression of target genes.

The primer sequences recommended for sequencing:
GUS promoter primer (4744-4762): 5’-CACGGGTTGGGGTTTCTAC-3’
M13 reverse primer (3338-3354): 5’-CAGGAAACAGCTATGAC-3’

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