6 research outputs found
A Generic System for the Expression and Purification of Soluble and Stable Influenza Neuraminidase
The influenza surface glycoprotein neuraminidase (NA) is essential for the efficient spread of the virus. Antiviral drugs such as Tamiflu (oseltamivir) and Relenza (zanamivir) that inhibit NA enzyme activity have been shown to be effective in the treatment of influenza infections. The recent βswine fluβ pandemic and world-wide emergence of Tamiflu-resistant seasonal human influenza A(H1N1) H274Y have highlighted the need for the ongoing development of new anti-virals, efficient production of vaccine proteins and novel diagnostic tools. Each of these goals could benefit from the production of large quantities of highly pure and stable NA. This publication describes a generic expression system for NAs in a baculovirus Expression Vector System (BEVS) that is capable of expressing milligram amounts of recombinant NA. To construct NAs with increased stability, the natural influenza NA stalk was replaced by two different artificial tetramerization domains that drive the formation of catalytically active NA homotetramers: GCN4-pLI from yeast or the Tetrabrachion tetramerization domain from Staphylothermus marinus. Both recombinant NAs are secreted as FLAG-tagged proteins to allow for rapid and simple purification. The Tetrabrachion-based NA showed good solubility, increased stability and biochemical properties closer to the original viral NA than the GCN4-pLI based construct. The expressed quantities and high quality of the purified recombinant NA suggest that this expression system is capable of producing recombinant NA for a broad range of applications including high-throughput drug screening, protein crystallisation, or vaccine development
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The human body at cellular resolution: the NIH Human Biomolecular Atlas Program
Abstract: Transformative technologies are enabling the construction of three-dimensional maps of tissues with unprecedented spatial and molecular resolution. Over the next seven years, the NIH Common Fund Human Biomolecular Atlas Program (HuBMAP) intends to develop a widely accessible framework for comprehensively mapping the human body at single-cell resolution by supporting technology development, data acquisition, and detailed spatial mapping. HuBMAP will integrate its efforts with other funding agencies, programs, consortia, and the biomedical research community at large towards the shared vision of a comprehensive, accessible three-dimensional molecular and cellular atlas of the human body, in health and under various disease conditions
Recommended from our members
The human body at cellular resolution: the NIH Human Biomolecular Atlas Program
Transformative technologies are enabling the construction of three
dimensional (3D) maps of tissues with unprecedented spatial and molecular
resolution. Over the next seven years, the NIH Common Fund Human Biomolecular
Atlas Program (HuBMAP) intends to develop a widely accessible framework for
comprehensively mapping the human body at single-cell resolution by supporting
technology development, data acquisition, and detailed spatial mapping. HuBMAP
will integrate its efforts with other funding agencies, programs, consortia,
and the biomedical research community at large towards the shared vision of a
comprehensive, accessible 3D molecular and cellular atlas of the human body, in
health and various disease settings
Recommended from our members
The human body at cellular resolution: the NIH Human Biomolecular Atlas Program
Transformative technologies are enabling the construction of three
dimensional (3D) maps of tissues with unprecedented spatial and molecular
resolution. Over the next seven years, the NIH Common Fund Human Biomolecular
Atlas Program (HuBMAP) intends to develop a widely accessible framework for
comprehensively mapping the human body at single-cell resolution by supporting
technology development, data acquisition, and detailed spatial mapping. HuBMAP
will integrate its efforts with other funding agencies, programs, consortia,
and the biomedical research community at large towards the shared vision of a
comprehensive, accessible 3D molecular and cellular atlas of the human body, in
health and various disease settings