Construction of engineered bacterium that binds to white spot syndrome virus
2014
Hochschulschrift
Zugriff:
102
White spot syndrome virus (WSSV) is a serious pathogen in shrimp culture, and there is no effective prevention method to resolve it so far. In order to reduce the concentration of WSSV in a breeding environment, this study aims to develop a bacterium that can bind with WSSV and remove it from water. Previous studies indicated that the WSSV Binding Protein (WBP) and Rab7 can bind to the VP26 and VP28 of WSSV, separately, in addition, the grouper Mx protein was also be demonstrated that Mx protein could bind with nervous necrosis virus (NNV) and against the infection of NNV. Therefore, we expressed WBP, Rab7 and Mx protein on bacterial surface to grenerate WSSV-binding bacterium and examine whether the bacterium can bind to the WSSV. The DNA fragments of WBP and Mx were amplified by assemble PCR, Rab7 gene was cloned by RT-PCR (Reverse Transcriptase PCR) and the RNA extracted from shrimp infected with WSSV.Subsequently, the virus binding genes were fused to C termini of lppompA or N-termini of AIDA, the Gram-negative surface display system, and formed lpp-WBP, lpp-Rab7, lpp-Mx, WBP-AIDA, Rab7-AIDA and Mx-AIDA, separatively. These recombinant plasmids were transformed into bacteria to generate lpp-WBP/BL21, lpp-Rab7/BL21, lpp-Mx/BL21, WBP-AIDA/BL21, Rab7-AIDA/BL21, and Mx-AIDA/BL21, separatively. Finally, we found that these bacteria can express protein on surface but failed to bind with WSSV by ELISA.
Titel: |
Construction of engineered bacterium that binds to white spot syndrome virus
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Autor/in / Beteiligte Person: | Ho, Cheng-Hung ; 何政鴻 |
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Veröffentlichung: | 2014 |
Medientyp: | Hochschulschrift |
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