Jumainah Emat (2014) Genetic transformation of glycine maxcallus using biolistic-method. Masters thesis, Universiti Malaysia Sabah.
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Abstract
Glycine max (Soybean) is cultivated globally and is a major source of plant protein. The agronomic traits include high growth rate and rapid maturity making it an ideal candidate plant for phytopharming of recombinant proteins. Interferon (Type I) is one of the components of the innate immune system, which in association with pattern recognition receptors, initiates response to viral pathogens. Currently, recombinant human interferon is produced in cell culture and bacteria. However, this process is not cost-effective. A gene construct encoding Human Type I interferon was inserted into the plant transformation vector pCAMBIA2301 for biolistic-mediated transformation into soybean callus. G. max callus, induced on Murashige & Skoog medium containing 2,4-D and Kinetin are at concentrations of 1+1 mg/L, 1+2 mg/L, 2+1 mg/L, 2+2 mg/L, and 2+4 mg/L. The best result of callus induction was at 1+2 mg/L, 2+1 mg/L, and 2+2 mg/L of 2,4-D combined with kinetin. Root differentiation from the callus occurred once the callus was transferred to medium containing a low concentration of sucrose in combination of NAA + BAP and IBA + BAP. About 55 % of the callus was succeeded to induce roots from 1+1 mg/L of NAA + BAP combination compared to only 32 % from 0.5+0.5 mg/L of concentration. Confirmation of the inserted plasmid DNA was done by detecting promoter gene using polymerase chain reaction (PCR), CaMV 35S gene, 298bp, from the plasmid and the GUS transient expression on the stained callus using X-gluc. The outcomes of this study will improve our understanding of the production of recombinant protein in agricultural crops especially soybean.
| Item Type: | Thesis (Masters) |
|---|---|
| Keyword: | Glycine max, Soybean tissue culture, Type I interferon, Phytopharming, Plant transformation |
| Subjects: | Q Science > QK Botany > QK1-989 Botany > QK710-899 Plant physiology |
| Department: | INSTITUTE > Biotechnology Research Institute (BRI) |
| Depositing User: | DG MASNIAH AHMAD - |
| Date Deposited: | 28 Apr 2025 15:53 |
| Last Modified: | 28 Apr 2025 15:53 |
| URI: | https://eprints.ums.edu.my/id/eprint/43590 |
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