규조류 Chaetoceros salsugineum 감염 nuclear inclusion virus (CsNIV)의 바이러스 유사입자 형성

Abstract

Virus-like particles (VLPs) are non-infectious and immunogenic, virus-mimicking protein assemblies (including host specific binding abilities). The research on this area increasing rapidly towards drug delivery system (DDS), lipoparticle technology and vaccine candidates. In the present study, the Chaetoceros salsuginium microalgal infecting Chaetoceros salsugineum nuclear inclusion virus (CsNIV) was used to produce VLP. The CsNIV is a 38nm icosahedral virus that replicates within the nucleus of C. salsugineum. The CsNIV capsid protein gene (1.2kb) was cloned into pEt 29b vector. The recombinant pET 29b-CsNIV plasmid was transformed into E.coli BL21(DE3). The CsNIV capsid protein was purified using Ni-NTA affinity column chromatography using elution buffer (20mM Tris-HCl, 300mM NaCl, 250mM imidazole, pH8.0). For further purification was performed in FPLC packed with Sephacryl S-500 resin. The purified VLPs was analysed using native-PAGE and the molecular weight of the VLP was found to be 46kDa. The CsNIV-VLPS was labelled with FITC by dissociation and reassociation process. The VPL and FITC combination was confirmed by 12% (w/v) SDS-PAGE and UV light. Further experiments need to find out the host specificity of novel algicidal compounds encapsulated with CsNIV-VLPs such as, HPLC analysis, FITC labelling, host specificity test against various diatoms. Therefore, we suggest that this study may help to produce an alternative, specific and highly effective bio-control tool to prevent the harmful algal bloom in marine waters.