Grasshoppers Regulate N: P Stoichiometric Homeostasis by Changing Phosphorus Contents in Their Frass
Viral protein U (Vpu) is a type-III integral membrane protein encoded by Human Immunodeficiency Virus-1 (HIV- 1). It is expressed in infected host cells and plays several roles in viral progeny escape from infected cells, including down-regulation of CD4 receptors. But key structure/function questions remain regarding the mechanisms by which the Vpu protein contributes to HIV-1 pathogenesis. Here we describe expression of Vpu in bacteria, its purification and characterization. We report the successful expression of PelB-Vpu in Escherichia coli using the leader peptide pectate lyase B (PelB) from Erwinia carotovora. The protein was detergent extractable and could be isolated in a very pure form. We demonstrate that the PelB signal peptide successfully targets Vpu to the cell membranes and inserts it as a type I membrane protein. PelB-Vpu was biophysically characterized by circular dichroism and dynamic light scattering experiments and was shown to be an excellent candidate for elucidating structural models.
Sources and strategies of finance studied and ultimately included in the tool were Loans, Equity, Membership, Crowdfunding, and Grants. The tool designed was a matrix that takes into account various criteria of the business (e.g. business lifecycle, organizational structure, business performance) and generates a financial plan based on these criteria and how they align with the selected business strategies. After strategies are found, stakeholders can search through an institutional database created in conjunction with the matrix tool to find possible institutional providers of financing that relate to the strategy or strategies found.
The tool has shown promise in identifying sources of finance for micro and small local food enterprises in practical use with hypothetical business cases, however further practical use is necessary to provide further input and revise the tool as needed. Ultimately, the tool will likely become fully user-friendly and stakeholders will not need the assistance of another expert helping them to use it. Finally, despite the promise of the tool itself, the fundamental and underlying problem that many of these businesses face (lack of infrastructure and knowledge) still exists, and while this tool can also help capacity-building efforts towards both those seeking and those providing finance, an institutional attitude adjustment towards social and alternative enterprises is necessary in order to further simplify the process of obtaining finance.