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- W581974545 abstract "The selectable marker system based on the Escherichia coli phosphomannoseisomerase (pmi) gene was adapted for genetic transformation of oil palm. This systemmakes use of the pmi gene that encodes phosphomannose isomerase, which convertsmannose-6-phosphate to fructose-6-phosphate and uses mannose as the selection agent.This is to anticipate the future requirement of using non-antibiotic resistance genes forthe commercialization of transgenic oil palm. The use of antibiotic or herbicide-basedselection systems has caused much public concern due to inadequate knowledge of theagents’ impact on the environment and on human health. In this study, fourtransformation vectors, namely pMI3, pMI11, pMI3G and pMI11G were constructedfor transforming oil palm. The pmi gene is driven by CaMV35S promoter in pMI3 andpMI3G, and Ubi1 promoter in pMI11 and pMI11G. gusA gene was also included in thepMI3G and pMI11G constructs. The gene constructs were transferred into oil palmembryogenic calli via biolistic-mediated transformation. Bombarded calli were selectedon medium supplemented with mannose as a carbon source. Results from kill curveexperiment indicated that oil palm embryogenic callus have little or no PMI activity andcannot utilize mannose as a carbon source. However, when calli were transformed witha pmi gene, the PMI activity was greatly increased and they could utilize mannoseefficiently as carbon source. For early identification of transgenic events, histochemicalstaining with 5-bromo-4-chloro-3-indolyl-s-D-glucuronide (X-Gluc) was used.Transgenic plants were confirmed by PCR and the transgene expression was detectedusing RT-PCR analysis. Transient expression results demonstrate that the Ubi1promoter is more efficient than the CaMV35S promoter in oil palm embryogenic calli.The insertion verification was confirmed with 98% homology observed to itscorresponding pmi gene from E. coli (Genbank accession no: M15380) via PCR directsequencing. In conclusion, the results of this study indicated that mannose selectionsystem can be used for oil palm transformation. Potentially this will make transgenic oilpalm acceptable in the future." @default.
- W581974545 created "2016-06-24" @default.
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- W581974545 date "2010-01-01" @default.
- W581974545 modified "2023-09-27" @default.
- W581974545 title "Biolistic transformation of oil palm using the phosphomannose isomerase (pmi) gene as the positive selectable marker / Bahariah Bohari" @default.
- W581974545 hasPublicationYear "2010" @default.
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