Construction of phosphomannose isomerase (PMI) transformation vectors and evaluation of the effectiveness of vectors in tobacco (Nicotiana tabacum L)
| Autor: | Bohari Bahariah, Ghulam Kadir Ahmad Parveez, Mat Yunus Abdul Masani, Norzulaani Khalid |
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| Rok vydání: | 2012 |
| Předmět: |
Genetics
Nicotiana tabacum fungi gusA Mannose General Medicine Genetically modified crops Biology Hypothesis biology.organism_classification phosphomannose isomerase tobacco Transformation (genetics) chemistry.chemical_compound Transformation vectors chemistry Restriction digest biolistics Cauliflower mosaic virus Energy source Gene |
| Zdroj: | Bioinformation |
| ISSN: | 0973-2063 |
| Popis: | Phosphomannose isomerase (pmi) gene isolated from Escherichia coli allows transgenic plants carrying it to convert mannose-6- phosphate (from mannose), a carbon source that could not be naturally utilized by plants into fructose-6-phosphate which can be utilized by plants as a carbon source. This conversion ability provides energy source to allow the transformed cells to survive on the medium containing mannose. In this study, four transformation vectors carrying the pmi gene alone or in combination with the β-glucuronidase (gusA) gene were constructed and driven by either the maize ubiquitin (Ubi1) or the cauliflower mosaic virus (CaMV35S) promoter. Restriction digestion, PCR amplification and sequencing were carried out to ensure sequence integrity and orientation. Tobacco was used as a model system to study the effectiveness of the constructs and selection system. PMI11G and pMI3G, which carry gusA gene, were used to study the gene transient expression in tobacco. PMI3 construct, which only carries the pmi gene driven by CaMV35S promoter, was stably transformed into tobacco using biolistics after selection on 30 g 1-1 mannose without sucrose. Transgenic plants were verified using PCR analysis. Abbreviations PMI/pmi - Phosphomannose isomerase, Ubi1 - Maize ubiquitin promoter, CaMV35S - Cauliflower mosaic virus 35S promoter, gusA - β-glucuronidase GUS reporter gene. |
| Databáze: | OpenAIRE |
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