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Replicon-based Japanese encephalitis virus vaccines elicit immune response in mice.

In this study, a replicon vaccine vector system for Japanese encephalitis virus (JEV) was established. The system included a trans-complementing cell line, a series of JEV DNA-based subgenomic replicons, and several encapsidated JEV propagation-deficient pseudoinfectious particles (PIPs). The DNA-based JEV replicon vectors, which deleted the structural coding region, could be able to self-replicate and express the reporter gene. A stable BHK packaging cell line named BHK-CME, which constitutively expressed the capsid protein C, the precursor membrane and envelope proteins (C-prM-E) of JEV, was…

An alphavirus replicon-derived candidate vaccine against Rift Valley fever virus.

Rift Valley fever virus (RVFV) is a mosquito-transmitted bunyavirus (genus Phlebovirus) associated with severe disease in livestock and fatal encephalitis or haemorrhagic fever in a proportion of infected humans. Although live attenuated and inactivated vaccines have been used in livestock, and on a limited scale in humans, there is a need for improved anti-RVFV vaccines. Towards this goal, Sindbis virus replicon vectors expressing the RVFV Gn and Gc glycoproteins, as well as the non-structural nsM protein, were constructed and evaluated for their ability to induce protective immune responses …

[Construction and application of subgenomic replicon vectors of Japanese encephalitis virus].

Based on the infectious clone of JEV vaccine SA14-14-2, the subgenomic replicons pCTCJEV, pCTMJEV with large deletions in the structural region were constructed. Then they were transfected into BHK-21 cell, the RNA replication of JEV subgenome can be monitored by RT-PCR and the non-structural protein can be found expressed in the cell by IFA. To explore the possibility of using a reporter gene assay to monitor synthesis of the positive-strand and the negative-strand JEV RNA, we inserted an enhanced green fluorescence protein (EGFP) gene into the 3'-UTR of pCTCJEV, pCTMJEV under the control of …

Construction of a Gordonia amarae shuttle vector using plasmid p1 replicon.

Members of the genus Gordonia are Gram-positive mycolic acid producing Actinomycetota, with important bioremediation capabilities, including hydrocarbon and heavy metal degradation. However, their hydrophobic cell surface also contributes to stabilisation of foams, making Gordonia species especially G. amarae, major causes of foaming in wastewater treatment plants worldwide. Genetic studies of Gordonia have been hindered by the lack of suitable plasmid vectors. In this study, a native Gordonia pseudoamarae plasmid, was used to construct a shuttle vector capable of replicating in Gordonia and E

An RNA replicon system to investigate promising inhibitors of feline coronavirus.

Feline infectious peritonitis (FIP) is a fatal feline disease, caused by a feline coronavirus (FCoV), namely feline infectious peritonitis virus (FIPV). We produced a baby hamster kidney 21 (BHK) cell line expressing a serotype I FCoV replicon RNA with a green fluorescent protein (GFP) reporter gene (BHK-F-Rep) and used it as an in vitro screening system to test different antiviral compounds. Two inhibitors of the FCoV main protease (Mpro), namely GC376 and Nirmatrelvir, as well as the nucleoside analog Remdesivir proved to be effective in inhibiting the replicon system. Different combinations…

Development of A MERS-CoV Replicon Cell Line for Antiviral Screening.

Middle East respiratory syndrome coronavirus (MERS-CoV) is the causative agent of a severe respiratory disease with a high mortality of ~ 35%. The lack of approved treatments for MERS-CoV infection underscores the need for a user-friendly system for rapid drug screening. In this study, we constructed a MERS-CoV replicon containing the Renilla luciferase (Rluc) reporter gene and a stable luciferase replicon-carrying cell line. Using this cell line, we showed that MERS-CoV replication was inhibited by combined application of lopinavir and ritonavir, indicating that this cell line can be used to …

Bivalent Junin & Machupo experimental vaccine based on alphavirus RNA replicon vector.

Junin (JUNV) and Machupo (MACV), two mammalian arenaviruses placed on the 2018 WHO watch list, are prevalent in South America causing Argent … s complicate approval of this vaccine in the US. To address these issues, we applied alphavirus RNA replicon vector technology based on the human Venezuelan equine encephalitis vaccine (VEEV) TC-83 to generate replication restricted virus-like-particles vectors (VLPVs) simultaneously expressing cellular glycoprotein precursors (GPC) of both viruses, JUNV a …

Potentiation of anthrax vaccines using protective antigen-expressing viral replicon vectors.

DNA vaccines require improvement for human use because they are generally weak stimulators of the immune system in humans. The efficacy of DNA vaccines can be improved using a viral replicon as vector to administer antigen of pathogen. In this study, we comprehensively evaluated the conventional non-viral DNA, viral replicon DNA or viral replicon particles (VRP) vaccines encoding different forms of anthrax protective antigen (PA) for specific immunity and protective potency against anthrax. Our current results clearly suggested that these viral replicon DNA or VRP vaccines derived from Semliki…

Replicon sequence typing of IncF plasmids carrying virulence and resistance determinants.

OBJECTIVES: IncF plasmids are frequently encountered in clinical enterobacterial strains associated with the dissemination of relevant antimicrobial resistance and virulence genes. These plasmids are usually heterogeneous in size and carry multiple replicons, and technical difficulties can impair the comparison and detection of related plasmids by restriction fragment length polymorphism analysis. We devised a rapid sequence-based typing scheme to categorize the members of this plasmid family into homogeneous groups. METHODS: We compared the available IncF replicon sequences, identifying the …

Анализ венесуэльских энцефалитных репликонных частиц, упакованных в различные пальто.

ВИЧ-репликоновая система венесуэльского энцефалита (VEE) была использована для производства подобных вирусу репликонных частиц (VRP), упакованных рядом различных гликопротеиновых халатов VEE (GP). Считается, что пальто GP отвечает за клеточный тропизм, отмеченный для VRP, и возможно, что различные пальто VEE GP могут иметь разные связи для клеток. Мы изучили VRP, упакованные в четыре различных пальто VEE GP, на предмет их способности инфицировать клетки в vitro и вызывать как юморальную, так и клеточную иммунную реакцию в vivo.МЕТОДОЛОГИЯ/ПРИНКИПАЛОВЫЕ ВЫВОДЫ: Подготовка ВРП была характерна дл

Совпадение в аннотации оригинала
BACKGROUND: The Venezuelan equine encephalitis (VEE) virus replicon system was used to produce virus-like replicon particles (VRP) packaged with a number of different VEE-derived glycoprotein (GP) coats. The GP coat is believed to be responsible for the cellular tropism noted for VRP and it is possible that different VEE GP coats may have different affinitie…

Development of a stable replicon system of a pangolin merbecovirus for antiviral screening.

• A replicon of the pangolin HKU4-related coronavirus (MjHKU4r-CoV-1) containing a nanoluciferase reporter gene was constructed. • A stable BHK21 cell line carrying the replicon has been generated. • This replicon system has been effectively employed in testing the antiviral activity of replication-targeting coronavirus drugs. • It offers a safe, efficient and convenient tool for screening antivirals against MjHKU4r-CoV-1 and other coronaviruses.

Development of an infectious clone and replicon system of norovirus GII.4.

Human norovirus (HuNoV) is one of the main causes of acute gastroenteritis worldwide and is responsible for at least 20% of all cases. The d … t protein (GFP) between the NTPase and p22 genes, also by Gibson Assembly, to construct a HuNoV-GFP replicon. Human Caco-2 cells were transfected with the full-length genomic clone and the replicon containing GFP. The gene encoding the VP1/VP2 capsid protein was expressed, which was indirect evidence of the synthesis of subgenomic RNAs and thus the negativ …

A pathogenicity island replicon in Staphylococcus aureus replicates as an unstable plasmid.

The SaPIs are 14- to 17-kb mobile pathogenicity islands in staphylococci that carry genes for superantigen toxins and other virulence factor … f the inducing phage. The ability of the SaPIs to replicate autonomously defines them as individual replicons and, like other prokaryotic replicons, they possess replicon-specific initiation functions. In this paper, we report identification of the SaPI replication origin (ori) and replication initiation protein (Rep), which has helicase as well as initiat …

Characterization of cell lines stably transfected with rubella virus replicons.

Rubella virus (RUBV) replicons expressing a drug resistance gene and a gene of interest were used to select cell lines uniformly harboring the replicon. Replicons expressing GFP and a virus capsid protein GFP fusion (C-GFP) were compared. Vero or BHK cells transfected with either replicon survived drug selection and grew into a monolayer. However, survival was ~9-fold greater following transfection with the C-GFP-replicon than with the GFP-expressing replicon and while the C-GFP-replicon cells grew similarly to non-transfected cells, the GFP-replicon cells grew slower. Neither was due to the a…

Multiple replicons constitute the 6.5-megabase genome of Burkholderia pseudomallei.

Burkholderia pseudomallei is a causative agent of melioidosis, a fatal tropical infectious disease endemic in Southeast Asia and Northern Au … ited in a public database. The results showed that the B. pseudomallei genome consists of two large replicons, and that both contain ribosomal RNA gene sequences, indicating the presence of two chromosomes. The classical arabinose-negative B. pseudomallei isolate K96243 has chromosomes of approximately 3563 +/- 73 and 2974 +/- 40 kilobase-pairs in size, gi …

Organization and replicon interactions within the highly segmented genome of Borrelia burgdorferi.

Borrelia burgdorferi, a causative agent of Lyme disease, contains the most segmented bacterial genome known to date, with one linear chromosome and over twenty plasmids. How this unusually complex genome is organized, and whether and how the different replicons interact are unclear. We recently demonstrated that B. burgdorferi is polyploid and that the copies of the chromosome and plasmids are regularly spaced in each cell, which is critical for faithful segregation of the genome to daughter cells. Regular spacing of the chromosome is controlled by two separate partitioning systems that involv…

A Yellow Fever 17D Virus Replicon-Based Vaccine Platform for Emerging Coronaviruses.

The tremendous global impact of the current SARS-CoV-2 pandemic, as well as other current and recent outbreaks of (re)emerging viruses, emphasize the need for fast-track development of effective vaccines. Yellow fever virus 17D (YF17D) is a live-attenuated virus vaccine with an impressive efficacy record in humans, and therefore, it is a very attractive platform for the development of novel chimeric vaccines against various pathogens. In the present study, we generated a YF17D-based replicon vaccine platform by replacing the prM and E surface proteins of YF17D with antigenic subdomains from th…

Replicon typing of plasmids in environmental Achromobacter sp. producing quinolone-resistant determinants.

This study aimed to investigate the antimicrobial resistance profile to quinolones, the presence of quinolone-resistant determinants and the plasmid replicon typing in environmental Achromobacter sp. isolated from Brazil. Soil and water samples were used for bacterial isolation. The antimicrobial susceptibility testing was performed by minimum inhibitory concentration method. The detection of mutations in the quinolone resistance-determining regions (QRDR) genes, the presence of plasmid-mediated quinolone resistance (PMQR) genes, and plasmid replicons were performed by PCR. A total of 16 isola…

Selection and Characterization of Rupintrivir-Resistant Norwalk Virus Replicon Cells In Vitro.

Human norovirus (HuNoV) is a major cause of nonbacterial gastroenteritis worldwide, yet despite its impact on society, vaccines and antivirals are currently lacking. A HuNoV replicon system has been widely applied to the evaluation of antiviral compounds and has thus accelerated the process of drug discovery against HuNoV infection. Rupintrivir, an irreversible inhibitor of the human rhinovirus 3C protease, has been reported to inhibit the replication of the Norwalk virus replicon via the inhibition of the norovirus protease. Here we report, for the first time, the generation of rupintrivir-re…

Replicon typing of plasmids in Escherichia coli producing extended-spectrum beta-lactamases.

OBJECTIVES: Escherichia coli producing CTX-M-15 and CTX-M-14 extended-spectrum beta-lactamases (ESBLs) are spreading worldwide. The aim of this work was to investigate the replicons involved in the emergence and spread of ESBLs in relation to ESBL type. METHODS: A collection of 125 TEM, SHV and CTX-M ESBL-producing E. coli strains was analysed. The replicons carrying the ESBLs and the total plasmid content of the strains have been characterized by PCR replicon typing in relation to the type of ESBL. The ESBL replicons were then compared with the replicon content of E. coli strains carrying TE…