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In vitro assembly of Penaeus monodon densovirus ( Pm DNV )‐like particles produced in a prokaryote expression system
Author(s) -
Sinnuengg Rapee,
Attasart Pongsopee,
Smith Duncan R,
Panyim Sakol,
Assavalapsakul Wanchai
Publication year - 2017
Publication title -
aquaculture research
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.646
H-Index - 89
eISSN - 1365-2109
pISSN - 1355-557X
DOI - 10.1111/are.13315
Subject(s) - penaeus monodon , biology , capsid , shrimp , white spot syndrome , virology , affinity chromatography , nucleic acid , virus , recombinant dna , rna silencing , rna , microbiology and biotechnology , rna interference , biochemistry , gene , fishery , enzyme
Viral diseases are a significant problem in the shrimp aquaculture industry as outbreaks can cause significant mortality and economic loss. While it has been shown that triggering the shrimp RNA interference pathway through ds RNA is a potentially viable treatment pathway, this approach is hampered by the lack of a suitable delivery mechanism. Virus‐like particles ( VLP s), which are structurally similar to native viruses but lack the genetic material, could possibly be developed as a delivery vehicle. To generate a candidate VLP , the Penaeus monodon densovirus ( Pm DNV ) capsid protein was cloned with an added histidine tag and expressed in an E. coli expression system. While the protein was expressed in inclusion bodies, the recombinant Pm DNV capsid protein could be dissolved and subsequently purified by nickel affinity column chromatography. The formation of VLP from this purified r P m DNV capsid protein was investigated by transmission electron microscopy, and Pm DNV ‐ VLP s were observed that looked similar to the native Pm DNV virion. Our results suggest that the Pm DNV ‐like particle could be promisingly applied towards vaccination and that this Pm DNV ‐like particle can potentially serve as a system for delivery of nucleic acids to trigger innate immunity in shrimp.