Study on the Interactions of Penaeus Monodon Laminin Receptor and White Spot Syndrome Virus Structural Proteins VP38B and VP52B

• Main Authors: HUANG, WEI-MING, 黃威閔
• Other Authors: CHANG, YUN-SHIANG
• Format: Others
• Language: zh-TW
• Published: 2017
• Online Access: http://ndltd.ncl.edu.tw/handle/49468950144713622450

碩士 === 大葉大學 === 分子生物科技學系碩士班 === 105 === White spot syndrome virus (WSSV) (family Nimaviridae, genus Whispovirus), originated in Fujian China in 1991. WSSV has a broad host range, infects most crustaceans, and causes high mortality. Effective method for complete treatment WSSV nfection caused diease is currently not available. A yeast two-hybrid screening results in our previous study identified interactions between the Penaeus monodon laminin receptor (PmLamr) and WSSV envelope proteins VP38B and VP52B. In the present study, these interactions were first confirmed by using co-immunoprecipitation assay. MBP-tagged recombinant VP38B (rMBP-VP38B) and VP52B (rMBP-VP52B) were produced using an Escherichia coli expression system, and their solubilities were determined. The purified rMBP-VP38B and rMBP-VP52B were then subjected to an enzyme-linked immunosorbent assay (ELISA) and a competitive ELISA to analyze their relationship with PmLamr during viral infection. The co-immunoprecipitation assays confirmed VP38B and VP52B could interact with PmLamr. In the protein solubility assays, rMBP-VP38B and rMBP-VP52B showed as water soluble proteins. The ELISA results showed that both rMBP-VP38B and rMBP-VP52B bound to His-tagged recombinant PmLamr (rHis-PmLamr) in a dose-dependent manner. The competitive ELISA revealed that the binding between rHis-PmLamr and rMBP-VP38B or rMBP-VP52B decreased with the increasing of WSSV concentration. The results revealed that VP38B and VP52B could bind to PmLamr and may play a role in the attachment of WSSV to the host cell. Data of the study could facilitate the understanding of the interaction between WSSV and PmLamr and would help in the development of antiviral agents.