Ali, Muhamad and DEPAMEDE, SULAIMAN N and Septyono, Bagus DH and Mukhlis, Alis and Amin, Muhamad and Ashari, Muhammad Turnitin: N-terminal Fusion of White Spot Syndrome Virus Envelope Protein VP28 with Glutathione-S-Transferase to Enhance Soluble Expression in Escherichia coli. Universitas Mataram.
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Abstract
The low production level and solubility of VP28 protein expressed in prokaryotic cells is considered to be a significant constraint for large-scale production of White Spot Syndrome Virus (WSSV) vaccine. Previous studies omitted N-terminal hydrophobic domain in VP28 gene in order to enhance the gene expression in E. coli. However, this approach generated incomplete VP28 protein, which may later affect the vaccine accuracy. Thus, this study has tried a protein fusion tag strategy in which the N-terminal hydrophobic domain of a full open reading frame (ORF) of VP28 gene was fused with a glutathione-S-transferase (GST) gene in a pGEX-4T-2 vector. The fused gene was subsequently expressed in E. coli BL21 (DE3) with 0.25 mM IPTG induction. The result showed significant increase in the expression level and solubility of the VP28 recombinant protein. In addition, the GST fusion facilitated the purification process of the protein using available commercial kit in down-stream steps for further analysis. These results suggested that the fusion technique is a potential approach to produce a complete VP28 protein for large-scale production of shrimp vaccine.
| Item Type: | Other |
|---|---|
| Keywords (Kata Kunci): | White Spot Syndrome Virus (WSSV), VP28, N-terminal hydrophobicdomain, GST-tag |
| Subjects: | S Agriculture > SF Animal culture |
| Divisions: | Fakultas Peternakan |
| Depositing User: | . Zulkarnain Zulkarnain |
| Date Deposited: | 11 Feb 2021 00:37 |
| Last Modified: | 11 Feb 2021 00:37 |
| URI: | http://eprints.unram.ac.id/id/eprint/20905 |
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