Publication: PRODUCTION OF EDIBLE HUMAN ROTAVIRUS VP6-SUBUNITS IN AGROBACTERIUM-MEDIATED TRANSFORMED CENTELLA ASIATICA CALLUS
| dc.contributor.author | WONG YING PEI | |
| dc.date.accessioned | 2023-10-06T15:40:56Z | |
| dc.date.available | 2023-10-06T15:40:56Z | |
| dc.date.issued | 2020 | |
| dc.description.abstract | Human rotaviruses (hRV) are one of the most common causes of severe diarrhoea in young children worldwide with high mortality rate especially in developing countries. Rotavirus VP6 protein has been suggested as a vaccine target due to its immunogenic and antigenic potential. Present study utilised the Agrobacterium-mediated transformation on Centella asiatica callus as the host to produce rotavirus VP6 subunit. The antibiotics - cefotaxime (250 mg/L), kanamycin (750 mg/L) and hygromycin (15 mg/L) were used in transformation process for transformant selection. Besides antibiotic susceptibility, bacterium concentrations, infection period and transformation method were also determined. The present study showed that Centella asiatica calli that were infected for45 minutes with Agrobacterium tumefaciens harbouring human rotavirus VP6genes (optical density6000.8) through needle puncture method exhibited a significant increment in its growth [5.4 ± 0.09 g fresh weight (FW) callus after four weeks of co-cultivation in antibiotic selection medium containing kanamycin and hygromycin. The transformation efficiency recorded under this optimal transformation protocols was 57.6%. The successful transformants were further confirmed using molecular techniques such as PCR(VP6gene has 1206bp)and western blot analysis(VP6 protein at 45 kDa). Quantification on the hRV-VP6 protein expressed in Centella asiatica callus recorded an amount of 0.16 to 0.46 mg/g FW callus. To the best of the knowledge, this study was the first report on the successful production of plant-based rotavirus VP6 subunit in Centella asiatica through Agrobacterium-mediated transformation. Thus, apart from adding value to the current applications of Centalla asistica callus, this study also serves as thenovel edible Centalla asiatica plants producing rotavirus vaccine that could potentially help in resolving the public health issues such as on the side effect of current available vaccines, anti-vaccine concerns and appropriate vaccination plan. | en_US |
| dc.identifier.uri | https://hdl.handle.net/20.500.14377/32344 | |
| dc.language.iso | en | en_US |
| dc.publisher | International Medical University | en_US |
| dc.subject | Rotavirus | en_US |
| dc.subject | Diarrhea | en_US |
| dc.subject | Child | en_US |
| dc.subject | Mortality | en_US |
| dc.subject | Agrobacterium | en_US |
| dc.subject | Centella | en_US |
| dc.subject | Anti-Bacterial Agents | en_US |
| dc.subject | Public Health | en_US |
| dc.subject | Vaccines | en_US |
| dc.title | PRODUCTION OF EDIBLE HUMAN ROTAVIRUS VP6-SUBUNITS IN AGROBACTERIUM-MEDIATED TRANSFORMED CENTELLA ASIATICA CALLUS | en_US |
| dc.type | Thesis | |
| dspace.entity.type | Publication |