Dragon grouper nervous necrosis virus (DGNNV), a betanodavirus, is the causative agent of viral nervous necrosis (VNN) in dragon grouper (Epinephelus lanceolatus). In our study, capsid protein of DGNNV was expressed in Escherichia coli. We mutated arginines at N-termini capsid protein to investigate the role of arginines at 29-31th position. When capsid protein lost 25 amino acids at N-termini VLPs form, mutation in any two arginines at 29-31 position to alanine could the prohibit VLPs formation. Another extending three arginines at 23-25 position wouldn¡¦t increase the RNA encapsulation into VLPs. Furthermore, N-termini mutated VLPs were all RNase resistance like wt-VLPs, but the yield was distinctly less than wt-VLPs. In the single point alanine mutations, the VLPs yield of R29A was apparently higher than others (R30A and R31A). Using circular dichroism to observe the thermal denature process and thermal stability of DGNNV VLPs, we found the Tm about 60¢J of VLPs wouldn¡¦t alter even if arginines at 23-31 position were mutated. The findings suggested the VLPs of mutated arginines at 23-31 position wouldn¡¦t affect RNase resistance and thermal stability, but the yield were lower. Another, the arginines at the 30 and 31 position is more important than at 29 position for formation of VLPs.
| Identifer | oai:union.ndltd.org:NSYSU/oai:NSYSU:etd-0122110-012836 |
| Date | 22 January 2010 |
| Creators | Huang, Xin-han |
| Contributors | Min-Ying Wang, Chan-Shing Lin, Chi-Hsin Hsu, Zhi-Hong Wen |
| Publisher | NSYSU |
| Source Sets | NSYSU Electronic Thesis and Dissertation Archive |
| Language | Cholon |
| Detected Language | English |
| Type | text |
| Format | application/pdf |
| Source | http://etd.lib.nsysu.edu.tw/ETD-db/ETD-search/view_etd?URN=etd-0122110-012836 |
| Rights | not_available, Copyright information available at source archive |
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