Abstract
Galectins are a conserved family of carbohydrate‐binding proteins involved in a wide variety of cellular processes, such as cell‐cell interactions, tumor proliferation and metastasis, regulation of innate and adaptive immune responses, and recognition of microbial pathogens. Based on distinctive structural features, galectins are classified into the “proto”, “chimera” and “tandem‐repeat” types. Our prior studies have shown that all three categories are expressed in zebrafish (Danio rerio), and suggested that DrGal1, DrGal3 and DrGal9 play a key role during experimental infection of zebrafish cell lines with infectious hematopoietic necrosis virus (IHNV). IHNV, an economically important pathogen of both wild and farmed salmonids, is a negative‐sense single‐stranded virus belonging to the genus Novirhabdovirus. The lack of standardized tools for the rigorous detection and quantification of IHNV viral particles in vitro, and commercial antibodies for use in standard immunoassays has seriously hindered progress toward the resolution of this problem. Thus, the development of tools and techniques that will enable the optimization of the IHNV‐zebrafish infection model for the mechanistic study of the IHNV infection process and the subsequent host immune response is of great interest. In this study, we designed a protocol for production and purification of the viral G (glycoprotein) protein of IHNV in recombinant bacteria, and initiated the production of polyclonal antibodies, that will enable us to visualize potential interactions between galectins and IHNV particles.Grant Funding Source: Supported by grant 5R01GM070589‐06 from the National Institutes of Health to GRV, and the Department of Biotechnology, New Delhi, India, to CR; NGM is supported by grant 5T32AI095190‐02; RK is supported by the ExPERT Program from the NSF
Published Version
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