Effects of copy number of an octopine synthase enhancer element and its distance from the TATA box on heterologous expression in transgenic tobacco
The relationship among motif copy number, its distance from the TATA box and expression level was analyzed in transformed tobacco plants. Single or multiple copies of octopine synthase (ocs) enhancer elements from the ocs promoter were linked to the minimal ocs promoter and a β-glucuronidase (GUS) reporter gene, then transformed stably into tobacco. Reporter gene assays revealed that mere repetition of the ocs enhancer sequence is not sufficient for promoter activity. Increasing the number of copies of the ocs element elevated the level of gene expression in an additive manner that was dependent of the element’s distance from the TATA box. To our knowledge, this is the first report in which the regulation of transgene expression by interactions between these two factors has been documented.
KeywordsSynthetic promoter Motif copy number Distance effect GUS activity
We thank Dr. Decai Cui for providing the construct 8. This research was supported by grants from the National Special Program for Research and Industrialization of Transgenic Plants (Grant No. J99-A-038) and the National Natural Science Foundation of China (Grant No. 30770145).
- Bhullar S, Chakravarthy S, Advani S et al (2003) Strategies for development of functionally equivalent promoters with minimum sequence homology for transgene expression in plants: cis-elements in a novel DNA context versus domain swapping. Plant Physiol 132:988–998. doi: 10.1104/pp.103.020602 PubMedCrossRefGoogle Scholar
- Geisler M, Kleczkowski LA, Karpinski S (2006) A universal algorithm for genome-wide in silicio identification of biologically significant gene promoter putative cis-regulatory-elements; identification of new elements for reactive oxygen species and sucrose signaling in Arabidopsis. Plant J 45(3):384–398. doi: 10.1111/j.1365-313X.2005.02634.x PubMedCrossRefGoogle Scholar
- Sawant SV, Kiran K, Mehrotra R et al (2005) A variety of synergistic and antagonistic interactions mediated by cis-acting DNA motifs regulate gene expression in plant cells and modulate stability of the transcription complex formed on a basal promoter. J Exp Bot 56:2345–2353. doi: 10.1093/jxb/eri227 PubMedCrossRefGoogle Scholar