A5081996486- Viral gastroenteritis research and epidemiology
- Animal Virus Infections Studies
- SARS-CoV-2 and COVID-19 Research
- Virus-based gene therapy research
- Mosquito-borne diseases and control
- Animal Disease Management and Epidemiology
- Viral Infections and Immunology Research
- Viral Infections and Vectors
- Insect symbiosis and bacterial influences
- Viral Infectious Diseases and Gene Expression in Insects
- Vector-Borne Animal Diseases
- Virology and Viral Diseases
- Studies on Chitinases and Chitosanases
- RNA Interference and Gene Delivery
- Monoclonal and Polyclonal Antibodies Research
- Bacteriophages and microbial interactions
- Plant and Fungal Interactions Research
- Plant Virus Research Studies
- Complement system in diseases
- Aerosol Filtration and Electrostatic Precipitation
- Human-Animal Interaction Studies
- Sirtuins and Resveratrol in Medicine
- T-cell and Retrovirus Studies
- T-cell and B-cell Immunology
- PARP inhibition in cancer therapy
Centro Nacional de Biotecnología
2013-2024
Centro Medico Nacional Siglo XXI
2024
Universidad Autónoma de Madrid
2000-2020
Consejo Superior de Investigaciones Científicas
1997-2015
Springer Publishing Company (United States)
2006
University of Iowa
2006
Iowa City Public Library
2006
In-Q-Tel
2003
University of Oxford
2001
Centro de Biología Molecular Severo Ochoa
1989-1999
A deletion mutant of severe acute respiratory syndrome coronavirus (SARS-CoV) has been engineered by deleting the structural E gene in an infectious cDNA clone that was constructed as a bacterial artificial chromosome (BAC). The recombinant virus lacking (rSARS-CoV-DeltaE) rescued Vero E6 cells. recovered grew E6, Huh-7, and CaCo-2 cells to titers 20-, 200-, 200-fold lower than wild-type virus, respectively, indicating although protein effect on growth, it is not essential for replication....
ABSTRACT Middle East respiratory syndrome coronavirus (MERS-CoV) is an emerging infecting humans that associated with acute pneumonia, occasional renal failure, and a high mortality rate considered threat to public health. The construction of full-length infectious cDNA clone the MERS-CoV genome in bacterial artificial chromosome reported here, providing reverse genetics system study molecular biology virus develop attenuated viruses as vaccine candidates. Following transfection clone, was...
The construction of cDNA clones encoding large-size RNA molecules biological interest, like coronavirus genomes, which are among the largest mature known to biology, has been hampered by instability those cDNAs in bacteria. Herein, we show that application two strategies, cloning into a bacterial artificial chromosome and nuclear expression RNAs typically produced within cytoplasm, is useful for engineering large molecules. A an infectious genome cloned as chromosome. rescued conserved all...
The engineering of a full-length infectious cDNA clone and functional replicon the severe acute respiratory syndrome coronavirus (SARS-CoV) Urbani strain as bacterial artificial chromosomes (BACs) is described in this study. In system, viral RNA was expressed cell nucleus under control cytomegalovirus promoter further amplified cytoplasm by replicase. Both were fully stable Escherichia coli. Using SARS-CoV replicon, we have shown that recently RNA-processing enzymes exoribonuclease,...
The construction of a set transmissible gastroenteritis coronavirus (TGEV)-derived replicons as bacterial artificial chromosomes is reported. These were generated by sequential deletion nonessential genes for virus replication, using modified TGEV full-length cDNA clone containing unique restriction sites between each pair consecutive genes. Efficient activity was associated with the presence nucleoprotein provided either in cis or trans. functional several cell lines, including human line...
Fractional killing is the main cause of tumour resistance to chemotherapy. This phenomenon observed even in genetically identical cancer cells homogeneous microenvironments. To understand this variable resistance, here we investigate individual responses TRAIL a clonal population HeLa using live-cell microscopy and computational modelling. We show that cellular mitochondrial content determines apoptotic fate modulates time death, with higher are more prone die. find all protein levels...
The pandemic coronavirus (CoV) disease 2019 (COVID-19) caused by severe acute respiratory syndrome CoV-2 (SARS-CoV-2) is a major threat to global human health. To date, there are no approved prophylactics or therapeutics available for COVID-19. Reverse genetics powerful approach understand factors involved in viral pathogenesis, antiviral screening, and vaccine development. In this study, we describe the feasibility of generating recombinant SARS-CoV-2 (rSARS-CoV-2) transfection single...
We have identified an open reading frame, EP402R, within the EcoRI E' fragment of African swine fever virus genome that encodes a polypeptide 402 amino acid residues homologous to adhesion receptor T cells, CD2. Transcription EP402R takes place during late phase replication. The disruption achieved through replacement 354-bp-long from by marker gene lacZ, does not affect growth rate in vitro but abrogates ability induce adsorption pig erythrocytes surface infected cells. This result...
The genome of African swine fever virus was screened for the existence repetitive sequences by hybridization between different cloned restriction fragments covering viral DNA. Several sets repeated were detected in located close to DNA ends. One these groups repetitions involved at both ends genome. remaining that exclusively left end. sequence a 3.2-kilobase segment spanning from 7.5 11 kilobases end, which showed complex pattern cross-hybridizations, determined. Two short and three long...
This study describes the assembly of a full-length cDNA clone human coronavirus (HCoV)-OC43 in bacterial artificial chromosome (BAC). The BAC containing infectious (pBAC-OC43(FL)) was assembled using two-part strategy. first step consisted introduction each end viral genome into with accessory sequences allowing proper transcription. second insertion whole HCoV-OC43 BAC. To produce recombinant particles, pBAC-OC43(FL) transfected BHK-21 cells. Recombinant virus displayed same phenotypic...
ABSTRACT The coronavirus (CoV) discontinuous transcription mechanism is driven by long-distance RNA-RNA interactions between transcription-regulating sequences (TRSs) located at the 5′ terminal leader (TRS-L) and also preceding each mRNA-coding sequence (TRS-B). contribution of host cell proteins to CoV needs additional information. Polypyrimidine tract-binding protein (PTB) was reproducibly identified in association with positive-sense RNAs transmissible gastroenteritis (TGEV) TRS-L TRS-B...
The full-length genome of the highly lethal feline infectious peritonitis virus (FIPV) strain DF-2 was sequenced and cloned into a bacterial artificial chromosome (BAC) to study role ORF3abc in FIPV-feline enteric coronavirus (FECV) transition. reverse genetic system allowed replacement truncated original FIPV with intact canine (CCoV) reference Elmo/02. vitro replication kinetics these two viruses studied CrFK FCWF-4 cell lines, as well peripheral blood monocytes. Both showed similar...
Abstract Here we report the rescue of a recombinant porcine reproductive and respiratory syndrome virus (PRRSV) carrying an enhanced green fluorescent protein (EGFP) reporter gene as separate transcription unit. A copy regulatory sequence for ORF6 (TRS6) was inserted between N 3′-UTR to drive EGFP yield general purpose expression vector. Successful recovery PRRSV obtained using RNA polymerase II promoter full-length genome, which assembled in bacterial artificial chromosome (BAC). The showed...
ABSTRACT A method to study the function of individual African swine fever virus (ASFV) gene products utilizing Escherichia coli lac repressor-operator system has been developed. Recombinant viruses containing both lacI encoding repressor and a strong late promoter modified by insertion one or two copies operator sequence at various positions were constructed. The ability each regulate expression firefly luciferase was assayed in presence absence inducer isopropyl β- d -thiogalactoside...
The recent outbreaks of Zika virus (ZIKV), its association with Guillain⁻Barré syndrome and fetal abnormalities, the lack approved vaccines antivirals, highlight importance developing countermeasures to combat ZIKV disease. In this respect, infectious clones constitute excellent tools accomplish these goals. However, flavivirus are often difficult work due toxicity some sequences in bacteria. To bypass problem, several alternative approaches have been applied for generation including, among...