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Tracking of the CaMV-35S promoter performance in GFP transgenic tobacco, with a special emphasis on flowers and reproductive organs, confirmed its predominant activity in vascular tissues
Authors:Marek Hra?ka  Slavomír Rakouský  Vladislav ?urn
Institution:(1) Faculty of Health and Social Studies, University of South Bohemia, Jírovcova 24, 370 01 Ceske Budejovice, Czech Republic;(2) Department of Genetics, Faculty of Science, University of South Bohemia, Branišovská 31, 370 05 Ceske Budejovice, Czech Republic;(3) Biotechnological Centre, Faculty of Agriculture, University of South Bohemia, Studentská 13, 370 05 Ceske Budejovice, Czech Republic
Abstract:Constitutive promoters are the most common promoters used to drive the expression of various genes in monocots and dicots. Therefore, it is of intense interest to ascertain their expression patterns in various plant species, organs and during their ontogenic development. In this study, the activity of the CaMV 35S promoter in transgenic tobacco plants was assessed. In contrast to other studies, performed rather on the primary transformants (T0 generation), here, individuals of T1 and T2 generations were used. The expression profiles of the CaMV 35S promoter were tracked within various plant organs and tissues using the GFP marker. Special attention was given to floral tissues for which the original data regarding the CaMV 35S expression were obtained. As expected, distinct developmental and organ/tissue specific expression patterns in a plant body were observed. CaMV 35S activity was detected in most of the plant tissues and during different developmental stages. The GFP signal was not visible in dry seeds only, but it became clearly apparent within 24–48 h after sowing onto the medium, what, among other things, enables the discrimination of transgenic and non-transgenic seeds/seedlings. Afterwards, the most pronounced GFP fluorescence intensity was usually visible in various vascular tissues of both, T1 and T2 plants, indicating the high promoter activity. A stable manifestation of the promoter was retained in the next T2 generation without any evident changes or losses of activity, showing the expression stability of the CaMV 35S.
Keywords:CaMV 35S promoter  Expression pattern  Fluorescence  Green fluorescent protein marker  Transgenic tobacco (Nicotiana tabacum)
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