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Expression of an Immunogenic Protein derived from Spike Protein of Porcine Epidemic Diarrhea Virus in two E. coli Strains |
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| รหัสดีโอไอ | |
| Title | Expression of an Immunogenic Protein derived from Spike Protein of Porcine Epidemic Diarrhea Virus in two E. coli Strains |
| Creator | Yaowaluck Maprang Roshorm |
| Contributor | Sochanwattey Meas, Pimploy Rattanaamnuaychai, Phenjun Mekvichitsaeng |
| Publisher | The Thai Society for Biotechnology |
| Publication Year | 2562 |
| Keyword | E. coli, PEDV, Protein Expression, Spike Protein, Vaccine |
| Abstract | Escherichia coli is recognized as a fast, easy and inexpensive host system for production of valuable proteins including subunit proteins for vaccination. In this study, we studied whether PEDVSME, a new immunogen designed from spike protein of PEDV, can be produced in E. coli by studying in two strains: BL21(DE3) and BL21(DE3)pLysS. The prokaryotic expression plasmid, pET28b, was generated to contain the PEDVSME gene. Recombinant clone of pET28b.PEDVSME was successfully generated and selected in E. coli DH5?. Two strains of E. coli, BL21(DE3) and BL21(DE3)pLysS, were then transformed with recombinant plasmid pET28b.PEDVSME. Expression of the PEDVSME gene was induced with 1 mM IPTG. Analyzed by SDS-PAGE and Western blot, PEDVSME was not detected in E. coli BL21(DE3) but highly expressed in strain BL21(DE3)pLysS. Importantly, although the PEDVSME protein could be observed in soluble form, majority of the protein was found in insoluble form (inclusion body). This work provides useful information regarding appropriate E. coli host strain for efficient production of the immunogen PEDVSME, bringing a feasibility of using PEDVSME as a subunit vaccine. |
| Language | EN |
| URL Website | http://tsb2019.com/ |
| Website title | The 31st Annual Meeting of the Thai Society for Biotechnology and International Conference (TSB 2019) |
