碩士 / 國立高雄海洋科技大學 / 海洋生物技術研究所 / 102 / We have performed the comparative proteomic research which based on the two-dimensional gel electrophoresis and mass spectrometry (2-DE/MS), it is found that the altered level of some identified proteins in shrimp, Litopenaeus vannamei, following a protein-based vaccination might be responsible for conferring resistance against White Spot Syndrome Virus (WSSV). In this study, after the proteomic analysis, the mRNA expression of six chosen proteins included Calreticulin (Crt), Ubiquitin (UB), Heat shock protein 70 (HSP70), Peroxiredoxin (Prx), Phosphopyruvate hydratase (PpH) and cytosolic Manganese superoxide dismutase (cytMnSOD) were compared between the vaccinated and PBS group, also in comparison with WSSV infected shrimp. Two WSSV envelope proteins, VP28 and VP36B, were expressed in Escherichia coli as subunit vaccines. Shrimp were intramuscularly injected with rVPs or PBS on day1 and day4, then on day5 till day7 gill and hemocyte tissues were sampled and assayed by real-time quantitative PCR. In the gill, expressions of Crt, UB and Prx mRNA significant increased for rVP28 group compared to control group. For both rVP28 and rVP36B groups the expression of HSP70 significant increased and the mRNA level of PpH was downregulated. In the hemocyte, expressions of Crt, UB, Prx and HSP70 mRNA significant increased for both rVP28 and rVP36B groups, the mRNA level of cytMnSOD was upregulated for rVP28 group only. After WSSV challenge, expressions of Crt, UB, Prx, HSP70 and PpH mRNA significant increased in gill, only the mRNA level of cytMnSOD was down regulated. We also cloned and sequenced the full length mRNA of two ubiquitin genes, UBS27 and UBL40 from Litopenaeus vannamei (Accession no. KJ831562 and KJ831563).
| Identifer | oai:union.ndltd.org:TW/102NKIM0270007 |
| Date | January 2014 |
| Creators | Shi-Wei Lin, 林世偉 |
| Contributors | Jyh-Ming Tsai, 蔡志明 |
| Source Sets | National Digital Library of Theses and Dissertations in Taiwan |
| Language | zh-TW |
| Detected Language | English |
| Type | 學位論文 ; thesis |
| Format | 68 |
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