AI Article Synopsis
- Recombinant DNA technology lets scientists insert specific genes into viruses to study how they work.
- This chapter focuses on using a virus called vaccinia that can help express a gene controlled by a special signal called the T7 promoter.
- They created a new virus that can turn on gene expression with a special chemical, making it easier to study how these genes behave.
Article Abstract
Recombinant DNA technology has made it possible to develop molecular cloning vectors that allow the expression of heterologous genes in a variety of animal viruses. This chapter discusses the use of vaccinia virus encoding bacteriophage T7 RNA polymerase as an expression vector system. A chosen gene is inserted into a plasmid vector designed to express genes under the control of the T7 promoter. Transient expression can then be achieved either by transfecting this plasmid into cells infected with the recombinant vaccinia virus expressing T7 RNA polymerase, vTF7-3 or by crossing this plasmid into the vaccinia virus genome and coinfecting cells with both viruses. Moreover, placement of lacO downstream of the vaccinia virus P11 late promoter regulating T7 RNA polymerase expression, and integration of lacI under vaccinia promoter control into the viral genome, vT7lacOI, yielded a recombinant virus capable of IPTG-inducible T7 promoter-controlled expression of foreign genes.
Download full-text PDF |
Source |
|---|---|
| http://dx.doi.org/10.1385/1-59259-789-0:041 | DOI Listing |
Publication Analysis
Top Keywords
Similar Publications
Domestic Dogs Exposed to Orthopoxvirus in Urban Areas of Brazil.
Viruses
January 2025
Laboratório de Vírus, Departamento de Microbiologia, Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antônio Carlos, 6627, Belo Horizonte 31270-901, Brazil.
Domestic animals can share viral pathogens with humans, acting mainly as a bridge host. The genus hosts important zoonotic species that have emerged in urban areas worldwide. Nevertheless, the role of companion animals, such as dogs and cats, in the circulation of orthopoxviruses in urban areas remains poorly understood.
View Article and Find Full Text PDFRepurposing Drugs for Synergistic Combination Therapies to Counteract Monkeypox Virus Tecovirimat Resistance.
Viruses
January 2025
Department of Immunology and Microbiology, The Scripps Research Institute, La Jolla, CA 92037, USA.
The ongoing monkeypox (mpox) disease outbreak has spread to multiple countries in Central Africa and evidence indicates it is driven by a more virulent clade I monkeypox virus (MPXV) strain than the clade II strain associated with the 2022 global mpox outbreak, which led the WHO to declare this mpox outbreak a public health emergency of international concern. The FDA-approved small molecule antiviral tecovirimat (TPOXX) is recommended to treat mpox cases with severe symptoms, but the limited efficacy of TPOXX and the emergence of TPOXX resistant MPXV variants has challenged this medical practice of care and highlighted the urgent need for alternative therapeutic strategies. In this study we have used vaccinia virus (VACV) as a surrogate of MPXV to assess the antiviral efficacy of combination therapy of TPOXX together with mycophenolate mofetil (MMF), an FDA-approved immunosuppressive agent that we have shown to inhibit VACV and MPXV, or the N-myristoyltransferase (NMT) inhibitor IMP-1088.
View Article and Find Full Text PDFeEF-2K Deficiency Boosts the Virus-Specific Effector CD8 T Cell Responses During Viral Infection.
Viruses
December 2024
Department of Microbial Pathogenesis and Immunology, Texas A&M University Health Science Center, Bryan, TX 77807, USA.
In this study, we revealed a critical role of eukaryotic elongation factor-2 kinase (eEF-2K), a negative regulator of protein synthesis, in regulating T cells during vaccinia virus (VACV) infection. We found that eEF-2K-deficient (eEF-2K⁻/⁻) mice exhibited a significantly higher proportion of VACV-specific effector CD8 T cells without compromising the development of VACV-specific memory CD8 T cells. RNA sequencing demonstrated that eEF-2K⁻/⁻ VACV-specific effector CD8 T cells had enhanced functionality, which improves their capacity to combat viral infection during the effector phase.
View Article and Find Full Text PDFGeographic Distribution of Vaccinia Virus, Diagnosis and Demographic Aspects of Affected Populations, Minas Gerais, Brazil, 2000-2023.
Viruses
December 2024
Instituto de Ciências Biológicas, Universidade Federal de Minas Gerais, Avenida Antônio Carlos, 6627, Belo Horizonte 31270-901, Brazil.
Since its first report in Brazil in 1999, outbreaks of exanthematous diseases caused by vaccinia virus (VACV) have been a recurring concern, particularly impacting rural regions. Minas Gerais (MG) State, Brazil, has emerged as the epicenter of bovine vaccinia (BV) outbreaks. This study presents a comprehensive overview of VACV circulation in MG State over the past two decades, examining the occurrence and distribution of poxvirus cases and outbreaks and the demographic characteristics of affected populations.
View Article and Find Full Text PDFSpecific Immune Responses and Oncolytic Effects Induced by EBV LMP2A-Armed Modified Ankara-Vaccinia Virus Vectored Vaccines in Nasopharyngeal Cancer.
Pharmaceutics
January 2025
NHC Key Laboratory of Systems Biology of Pathogens, Institute of Pathogen Biology, Chinese Academy of Medical Sciences, Peking Union Medical College, Beijing 100005, China.
Background: The Epstein-Barr virus (EBV) is intricately linked to a range of human malignancies, with EBV latent membrane protein 2A (LMP2A) emerging as a potential target antigen for immunotherapeutic strategies in the treatment of nasopharyngeal carcinoma (NPC).
Methods: The modified vaccinia virus Ankara (MVA) is universally used in vector vaccine research because of its excellent safety profile and highly efficient recombinant gene expression. Here, we constructed a novel MVA-LMP2A recombinant virus and investigated its specific immune response induction and oncolytic effect.
Want AI Summaries of new PubMed Abstracts delivered to your In-box?
Enter search terms and have AI summaries delivered each week - change queries or unsubscribe any time!