MeEF1?3 PROMOTER ACTIVITY IN VEGETATIVE ORGANS OF Nicotiana tabacum BY Agrobacterium-MEDIATED TRANSFORMATION
Promoter is a DNA sequences which regulate gene expression. MeEF1?3 is a member of EF1? family gene promoters isolated from cassava (Manihot esculenta Crantz.). Previous studies informed that this promoter were transiently active in roots, stems, leaves, and fruits of corn and tomato, also in Musa a...
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| Format: | Theses |
| Language: | Indonesia |
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| Online Access: | https://digilib.itb.ac.id/gdl/view/36183 |
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| Institution: | Institut Teknologi Bandung |
| Language: | Indonesia |
| Summary: | Promoter is a DNA sequences which regulate gene expression. MeEF1?3 is a member of EF1? family gene promoters isolated from cassava (Manihot esculenta Crantz.). Previous studies informed that this promoter were transiently active in roots, stems, leaves, and fruits of corn and tomato, also in Musa acuminata fruits and tobacco seedling. However, this expression could not be observed in tobacco seedling’s roots. This phenomenon indicates that MeEF1?3 promoter requires tissue specificity. MeEF1?3 promoter is a newly identified promoter which has not been characterized in stable expression. Therefore, stable analysis should be conducted to assess the precise expression pattern of MeEF1?3 promoter. The objective of this study is to evaluate MeEF1?3 promoter activity in various organs of Nicotiana tabacum Linn. as a result of Agrobacterium-mediated transformation. Construction of MeEF1?3 promoter which regulate GUS gene in pBI121 binary vector was introduced into tobacco explant. The result showed that kanamycin minimum concentration for selection medium was 100 mgL-1. Compare to the negative control (without binary vector), histochemical analysis of putative trangenic plantlet exhibited that GUS was regulated by MeEF1?3 promoter in stems and leaves (mainly in midrib), but not in roots. Analysis of stems and leaves section indicate that GUS was only expressed in phloem tissues, although the expression was qualitatively lower than GUS regulated by CaMV35S promoter (positive control). The absence of GUS expression in all root tissues describes that MeEF1?3 promoter was only active in tobacco aerial organs, specifically in phloem cells. Transformation efficiency for the method used in this study was 11,3%±4,2. |
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