Construction of a reverse genetic system for porcine astrovirus

2018 ◽  
Vol 163 (6) ◽  
pp. 1511-1518 ◽  
Author(s):  
Yifeng Qin ◽  
Qingli Fang ◽  
Huan Liu ◽  
Chengyuan Ji ◽  
Ying Chen ◽  
...  
Keyword(s):  
Genetic System ◽  
Reverse Genetic System ◽  
Reverse Genetic ◽  
Porcine Astrovirus

scholarly journals Development of a Reverse Genetic System to Generate Recombinant Chimeric Tacaribe Virus that Expresses Junín Virus Glycoproteins

Pathogens ◽  
2020 ◽  
Vol 9 (11) ◽  
pp. 948
Author(s):  
Sabrina Foscaldi ◽  
María Eugenia Loureiro ◽  
Claudia Sepúlveda ◽  
Carlos Palacios ◽  
María Belén Forlenza ◽  
...  
Keyword(s):  
Hemorrhagic Fever ◽  
Genetic System ◽  
Reverse Genetic System ◽  
Dependent Manner ◽  
Reverse Genetic ◽  
Junin Virus ◽  
Tacaribe Virus ◽  
Growth Properties ◽  
Intracellular Expression ◽  
Junín Virus

Mammarenaviruses are enveloped and segmented negative-stranded RNA viruses that comprise several pathogenic members associated with severe human hemorrhagic fevers. Tacaribe virus (TCRV) is the prototype for the New World group of mammarenaviruses and is not only naturally attenuated but also phylogenetically and antigenically related to all South American pathogenic mammarenaviruses, particularly the Junín virus (JUNV), which is the etiological agent of Argentinian hemorrhagic fever (AHF). Moreover, since TCRV protects guinea pigs and non-human primates from lethal challenges with pathogenic strains of JUNV, it has already been considered as a potential live-attenuated virus vaccine candidate against AHF. Here, we report the development of a reverse genetic system that relies on T7 polymerase-driven intracellular expression of the complementary copy (antigenome) of both viral S and L RNA segments. Using this approach, we successfully recovered recombinant TCRV (rTCRV) that displayed growth properties resembling those of authentic TCRV. We also generated a chimeric recombinant TCRV expressing the JUNV glycoproteins, which propagated similarly to wild-type rTCRV. Moreover, a controlled modification within the S RNA 5′ non-coding terminal sequence diminished rTCRV propagation in a cell-type dependent manner, giving rise to new perspectives where the incorporation of additional attenuation markers could contribute to develop safe rTCRV-based vaccines against pathogenic mammarenaviruses.

scholarly journals Establishment of an efficient reverse genetic system of Mumps virus S79 from cloned DNA

2019 ◽  
Vol 15 (5) ◽  
pp. 499-505 ◽  
Author(s):  
Duo Zhou ◽  
Meng-Ying Zhu ◽  
Yi-Long Wang ◽  
Xiao-Qiang Hao ◽  
Dong-Ming Zhou ◽  
...  
Keyword(s):  
Genetic System ◽  
Mumps Virus ◽  
Reverse Genetic System ◽  
Reverse Genetic

scholarly journals Characterization and reverse genetic establishment of cattle derived Akabane virus in China

2021 ◽  
Vol 17 (1) ◽  
Author(s):  
Dongjie Chen ◽  
Di Wang ◽  
Fang Wei ◽  
Yufang Kong ◽  
Junhua Deng ◽  
...  
Keyword(s):  
Genetic System ◽  
Reverse Genetic System ◽  
Akabane Virus ◽  
Reverse Genetic ◽  
Wild Type ◽  
T7 Promoter ◽  
Cytopathic Effects ◽  
The World ◽  
L Segment ◽  
Genome Manipulation

Abstract Background Akabane virus (AKAV) is an important insect-borne virus which is widely distributed throughout the world except the Europe and is considered as a great threat to herbivore health. Results An AKAV strain defined as TJ2016 was firstly isolated from the bovine sera in China in 2016. Sequence analysis of the S and M segments suggested that the isolated AKAV strain was closely related to the AKAV strains JaGAr39 and JaLAB39, which belonged to AKAV genogroup II. To further study the pathogenic mechanism of AKAV, the full-length cDNA clone of TJ2016 S, M, and L segment was constructed separately into the TVT7R plasmid at the downsteam of T7 promoter and named as TVT7R-S, TVT7R-M, and TVT7R-L, respectively. The above three plasmids were further transfected into the BSR-T7/5 cells simultaneously with a ratio of 1:1:1 to produce the rescued virus AKAV. Compared with the parental wild type AKAV (wtAKAV), the rescued virus (rAKAV) was proved to be with similar cytopathic effects (CPE), plaque sizes and growth kinetics in BHK-21 cells. Conclusion We successfully isolated a AKAV strain TJ2016 from the sera of cattle and established a reverse genetic platform for AKAV genome manipulation. The established reverse genetic system is also a powerful tool for further research on AKAV pathogenesis and even vaccine studies.

scholarly journals Development of a Reverse Genetic System for Infectious Salmon Anemia Virus: Rescue of Recombinant Fluorescent Virus by Using Salmon Internal Transcribed Spacer Region 1 as a Novel Promoter

2014 ◽  
Vol 81 (4) ◽  
pp. 1210-1224 ◽  
Author(s):  
Daniela Toro-Ascuy ◽  
Carolina Tambley ◽  
Carolina Beltran ◽  
Carolina Mascayano ◽  
Nicolas Sandoval ◽  
...  
Keyword(s):  
Atlantic Salmon ◽  
Cytopathic Effect ◽  
Genetic System ◽  
Reverse Genetic System ◽  
Reverse Genetic ◽  
Wild Type ◽  
Internal Transcribed Spacer Region ◽  
Content Type ◽  
Recombinant Viruses ◽  
Infectious Salmon Anemia

ABSTRACTInfectious salmon anemia (ISA) is a serious disease of marine-farmed Atlantic salmon (Salmo salar) caused by ISA virus (ISAV), belonging to the genusIsavirus, familyOrthomyxoviridae. There is an urgent need to understand the virulence factors and pathogenic mechanisms of ISAV and to develop new vaccine approaches. Using a recombinant molecular biology approach, we report the development of a plasmid-based reverse genetic system for ISAV, which includes the use of a novel fish promoter, the Atlantic salmon internal transcribed spacer region 1 (ITS-1). Salmon cells cotransfected with pSS-URG-based vectors expressing the eight viral RNA segments and four cytomegalovirus (CMV)-based vectors that express the four proteins of the ISAV ribonucleoprotein complex allowed the generation of infectious recombinant ISAV (rISAV). We generated three recombinant viruses, wild-type rISAV901_09and rISAVrS6-NotI-HPRcontaining a NotI restriction site and rISAVS6/EGFP-HPRharboring the open reading frame of enhanced green fluorescent protein (EGFP), both within the highly polymorphic region (HPR) of segment 6. All rescued viruses showed replication activity and cytopathic effect in Atlantic salmon kidney-infected cells. The fluorescent recombinant viruses also showed a characteristic cytopathic effect in salmon cells, and the viruses replicated to a titer of 6.5 × 105PFU/ml, similar to that of the wild-type virus. This novel reverse genetics system offers a powerful tool to study the molecular biology of ISAV and to develop a new generation of ISAV vaccines to prevent and mitigate ISAV infection, which has had a profound effect on the salmon industry.

Production of infectious dromedary camel hepatitis E virus by a reverse genetic system: Potential for zoonotic infection

2016 ◽  
Vol 65 (6) ◽  
pp. 1104-1111 ◽  
Author(s):  
Tian-Cheng Li ◽  
Xianfeng Zhou ◽  
Sayaka Yoshizaki ◽  
Yasushi Ami ◽  
Yuriko Suzaki ◽  
...  
Keyword(s):  
Hepatitis E Virus ◽  
Genetic System ◽  
Hepatitis E ◽  
Reverse Genetic System ◽  
Dromedary Camel ◽  
Reverse Genetic ◽  
Zoonotic Infection

scholarly journals Development and Characterization of a Reverse Genetic System for Studying Dengue Virus Serotype 3 Strain Variation and Neutralization

2012 ◽  
Vol 6 (2) ◽  
pp. e1486 ◽  
Author(s):  
William B. Messer ◽  
Boyd Yount ◽  
Kari E. Hacker ◽  
Eric F. Donaldson ◽  
Jeremy P. Huynh ◽  
...  
Keyword(s):  
Dengue Virus ◽  
Genetic System ◽  
Reverse Genetic System ◽  
Reverse Genetic ◽  
Strain Variation ◽  
Virus Serotype

scholarly journals Recombinant Mouse Hepatitis Virus Strain A59 from Cloned, Full-Length cDNA Replicates to High Titers In Vitro and Is Fully Pathogenic In Vivo

2005 ◽  
Vol 79 (5) ◽  
pp. 3097-3106 ◽  
Author(s):  
Scott E. Coley ◽  
Ehud Lavi ◽  
Stanley G. Sawicki ◽  
Li Fu ◽  
Barbara Schelle ◽  
...  
Keyword(s):  
Vaccinia Virus ◽  
Hepatitis Virus ◽  
Mouse Hepatitis Virus ◽  
Genetic System ◽  
Full Length ◽  
Reverse Genetic System ◽  
Plaque Morphology ◽  
Reverse Genetic ◽  
Group Ii ◽  
Cloned Cdna

ABSTRACT Mouse hepatitis virus (MHV) is the prototype of group II coronaviruses and one of the most extensively studied coronaviruses. Here, we describe a reverse genetic system for MHV (strain A59) based upon the cloning of a full-length genomic cDNA in vaccinia virus. We show that the recombinant virus generated from cloned cDNA replicates to the same titers as the parental virus in cell culture (∼109 PFU/ml), has the same plaque morphology, and produces the same amounts and proportions of genomic and subgenomic mRNAs in virus-infected cells. In a mouse model of neurological infection, the recombinant and parental viruses are equally virulent, they replicate to the same titers in brain and liver, and they induce similar patterns of acute hepatitis, acute meningoencephalitis, and chronic demyelination. We also describe improvements in the use of the coronavirus reverse genetic system based on vaccinia virus cloning vectors. These modifications facilitate (i) the mutagenesis of cloned cDNA by using vaccinia virus-mediated homologous recombination and (ii) the rescue of recombinant coronaviruses by using a stable nucleocapsid protein-expressing cell line for the electroporation of infectious full-length genomes. Thus, our system represents a versatile and universal tool to study all aspects of MHV molecular biology and pathogenesis. We expect this system to provide valuable insights into the replication of group II coronaviruses that may lead to the development of novel strategies against coronavirus infections, including the related severe acute respiratory syndrome coronavirus.

Sign in / Sign up

Export Citation Format

Share Document