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Radovica-Spalvina, I.

Publications and source records attributed to Radovica-Spalvina, I..

2 recordsLinked to original sources

Investigation of plant virus-like particle formation in bacterial and yeast expression systems

Virus-like particles (VLPs) have garnered significant attention due to their potential applications in various fields, particularly in biomedicine, where they are used for drug delivery, vaccine development, and as diagnostic tools. Plant virus-derived VLPs have been especially successful, leading to a surge in research aimed at developing new VLPs. Despite this growing interest, not all attempts at VLP formation are successful. Understanding the factors that contribute to successful VLP assembly is crucial for advancing this field. In this study, we focus on the coat proteins (CPs) of three plant viruses belonging to the Sobemovirus genus as a model to investigate the process of VLP formation. Our findings demonstrated that the strong binding of CPs to ssDNA can be a major reason for unsuccessful VLP production. This issue can potentially be overcome by transitioning from an episomal to a chromosome-integrated expression system. By doing so, we aim to elucidate the mechanisms that facilitate successful VLP assembly. Our findings provide valuable insights into the properties of sobemovirus CPs and highlight their potential for encapsulating foreign nucleic acids, thereby providing additional applications as nanocontainers or vaccine platforms with interior modification capabilities for enhanced immune response. Moreover, our RNA-Seq data indicates that sobemovirus-derived VLPs predominantly package their CP mRNA, irrespective of the expression system used. This characteristic can be utilized for the functional encapsulation of nucleic acids, enhancing the versatility and utility of these VLPs in various biotechnological applications.

molecular biology↗

Ryegrass mottle virus complete genome determination and development of infectious cDNA by combining two methods - 3' RACE and 5' RACE-seq

Sobemovirus ryegrass mottle virus (RGMoV) is a single-stranded positive virus with a 30 nm viral particle size. It exhibits T=3 symmetry, with 180 coat protein (CP) subunits forming the virus structure. The RGMoV genome comprises five open reading frames, encoding P1, Px, a membrane-anchored 3C-like serine protease, a virus genome-linked protein, P16, an RNA-dependent RNA polymerase, and a coat protein. The RGMoV genome size varies, ranging from 4175 nt (MW411579.1) to 4253 nt (MW411579.1) in deposited sequences. An earlier deposited RGMoV complete genome sequence of 4212 nt length (EF091714.1) was utilized to develop an infectious complementary DNA (icDNA) construct for in vitro gRNA transcription from the T7 promoter. However, when the transcribed gRNA was introduced to oat plants, it failed to induce viral infection. This indicated the potential absence of certain sequences in either the 5 or 3 untranslated regions (UTR) or both. To resolve this, the complete sequence of the 3 UTR was determined through 3 end RACE, while the 5 UTR was identified using high-throughput sequencing (HTS) - 5 RACE-seq. Only the icDNA vector containing both newly identified UTR sequences proved infectious, resulting in classical viral infection symptoms and subsequent propagation of progeny viruses, exhibiting the ability to cause repeated infection in oat plants after at least one passage. The successful generation of the icDNA highlights the synergistic potential of utilizing both methods when one approach alone fails. Furthermore, this study demonstrates the reliability of HTS as a method for determining the complete genome sequence of viral genomes.

molecular biology↗