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Agrobacterium-Mediated Virus-Induced Gene Silencing Assay In Cotton
Published on: August 20, 2011
Functional characterization of a bidirectional plant promoter from cotton leaf curl Burewala virus using an
Muhammad Aleem Ashraf1, Ahmad Ali Shahid2, Abdul Qayyum Rao3
1Plant Biotechnology Laboratory, Centre of Excellence in Molecular Biology (CEMB), University of the Punjab, Lahore 53700, Pakistan. muhammad.aleem@cemb.edu.pk.
Abstract:
The C1 promoter expressing the AC1 gene, and V1 promoter expressing the AV1 gene are located in opposite orientations in the large intergenic region of the Cotton leaf curl Burewala virus (CLCuBuV) genome. Agro-infiltration was used to transiently express putative promoter constructs in Nicotiana tabacum and Gossypium hirsutum leaves, which was monitored by a GUS reporter gene, and revealed that the bidirectional promoter of CLCuBuV transcriptionally regulates both the AC1 and AV1 genes. The CLCuBuV C1 gene promoter showed a strong, consistent transient expression of the reporter gene (GUS) in N. tabacum and G. hirsutum leaves and exhibited GUS activity two- to three-fold higher than the CaMV 35S promoter. The CLCuBuV bidirectional gene promoter is a nearly constitutive promoter that contains basic conserved elements. Many cis-regulatory elements (CREs) were also analyzed within the bidirectional plant promoters of CLCuBuV and closely related geminiviruses, which may be helpful in understanding the transcriptional regulation of both the virus and host plant.

