Please use this identifier to cite or link to this item:
https://ptsldigital.ukm.my/jspui/handle/123456789/500477Full metadata record
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.advisor | Ismanizan Ismail, Prof. Madya Dr. | |
| dc.contributor.author | Rima Saed Mustafa Taha (P39149) | |
| dc.date.accessioned | 2023-10-13T09:44:08Z | - |
| dc.date.available | 2023-10-13T09:44:08Z | - |
| dc.date.issued | 2012-05-24 | |
| dc.identifier.other | ukmvital:120754 | |
| dc.identifier.uri | https://ptsldigital.ukm.my/jspui/handle/123456789/500477 | - |
| dc.description | In this study the oil palm stearoyl-ACP desaturase gene promoter (Des) specificity and strength were evaluated and compared to cauliflower mosaic virus CaMV35S promoter in transgenic tomato. In order to carry out the functional analysis of the Des promoter on the expression levels in mesocarp tissue, and to define the minimal functional Des promoter; chimeric fusions between 314, 600, 812, 919, and 1032 bp of the 5' flanking region of the stearoyl-ACP desaturase gene and the β-glucuronidase (gus) coding sequence were subcloned into pCAMBIA 1301 vectors. An efficient protocol for Agrobacterium-mediated transformation and regeneration of local tomato MTI variety using hygromycin as a selective agent was developed. Hygromycin with the concentration of 4 mg/L and 8 mg/L were used as selective agent for callus induction in cotyledon and hypocotyls explants respectively for the first month. The concentration of hygromycin was increased to 15 mg/L in the second month, and subsequently reduced to 10 mg/L for root induction, resulting in the regeneration of stable transgenic plants carrying CaMV35S and Des promoters. The integration of the transgene into the tomato genome was confirmed by PCR amplification and Southern hybridization. The results for histochemical GUS staining in fruits, seeds and vegetative tissues of mature T0 and T1 transgenic tomato plants showed that the full length of Des promoter drove the expression of the gus gene in the fruit tissues and seeds. As expected, seeds, vegetative and fruit tissues of transgenic plants carrying the gus gene under the control of CaMV35S showed different levels of GUS activity. Analysis of the deletion series of Des promoter showed the gus gene expression in tomato fruit tissues only and no expression was observed in the vegetative tissues of the transgenic plants. The highest GUS activity was detected in the fruit tissue (vascular tissue, septa, endocarp, mesocarp and columella) and seeds which carried the promoter region located between -590 to +10 bp. This suggests the possible presence of negative regulatory region between -590 and -909 and that the 600 bp of Des promoter contains all cis-elements needed for expression in fruit and seed. Promoter deletion analyses have shown that the shortest promoter region (Des5, 314 bp) represents the minimum promoter region which directed considerably low level of GUS activity in columella of the tomato fruits and seeds. The quantitative analysis of GUS activity by flourometric assay has demonstrated 4-fold higher activity of the Des1 promoter compared to CaMV35S promoter. However, the strongest gus expression was driven by Des 4 promoter (1637.6 pmol/μg/min). Semi quantitative RT-PCR and Northern hybridization analyses have further supported the highest gus expression in fruits driven by Des 4. These results demonstrate the potential of the Des promoter as a tool for genetic manipulation of oil palm and other dicot transformation systems for the manipulation of the nutritional value and quality of fruits by genetic engineering.,Certification of Masters/ Doctorial Thesis" is not available | |
| dc.language.iso | eng | |
| dc.publisher | UKM, Bangi | |
| dc.relation | Faculty of Science and Technology / Fakulti Sains dan Teknologi | |
| dc.rights | UKM | |
| dc.subject | Dissertations, Academic -- Malaysia | |
| dc.subject | Promoters (Genetics) | |
| dc.subject | Agricultural chemicals | |
| dc.subject | Molecular genetics | |
| dc.subject | Transgenic plants-Plant molecular genetics | |
| dc.title | Molecular dissection of the mesocarp-specific Stearoyl-ACP desaturase gene promoter of oil palm and ots functional analysis in transgenic tomato | |
| dc.type | Theses | |
| dc.format.pages | 161 | |
| dc.identifier.callno | QK981.4.T337 2012 tesis | |
| dc.identifier.barcode | 002714 (2012) | |
| Appears in Collections: | Faculty of Science and Technology / Fakulti Sains dan Teknologi | |
Files in This Item:
| File | Description | Size | Format | |
|---|---|---|---|---|
| ukmvital_120754+SOURCE1+SOURCE1.0.PDF Restricted Access | 1.32 MB | Adobe PDF |  View/Open |
Items in DSpace are protected by copyright, with all rights reserved, unless otherwise indicated.