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Gary Blissard
 &emdash;  Professor

Gary Blissard
Office/Lab: 201/202
Affiliations:
  • Department of Microbiology & Immunology
  • Department of Entomology
  • Cornell University
Graduate Fields: Entomology; Comparative Biomedical Sciences
Research Areas:

Publications

  • Multifaceted biological insights from a draft genome sequence of the tobacco hornworm moth, Manduca sexta 2016

    Kanost, M.R., Arrese, E.L., Cao, X., Chen, Y.R., Chellapilla, S., Goldsmith, M.R., Grosse-Wilde, E., Heckel, D.G., Herndon, N., Jiang, H., et al
    Insect Biochem. Mol. Bio. 76,  118-147
    Full text...
  • Trichoplusia ni Kinesin-1 associates with AcMNPV nucleocapsid proteins and is required for the production of budded virus 2016

    Biswas, S., G. W. Blissard, and D. A. Theilmann
    Journal of Virology 90,  3480-3495
    Full text...
  • Autographa californica multiple nucleopolyhedrovirus GP64 protein: Analysis of domain I and V amino acid interactions and membrane fusion activity 2015

    Yu, Q., Blissard, G.W., Liu, T.X., Li, Z.
    Virology 488,  259-270
    Full text...
  • The immune signaling pathways of Manduca sexta 2015

    Cao, X., Y. He, Y. Hu, Y. Wang, Y. R. Chen, B. Bryant, R. J. Clem, L. M. Schwartz, G. Blissard, and H. Jiang
    Insect Biochem. Mol. Biol. 62,  64-74
    Full text...
  • Sequence conservation, phylogenetic relationships, and expression profiles of nondigestive serine proteases and serine protease homologs in Manduca sexta 2015

    Cao, X., He, Y., Hu, Y., Zhang, X., Wang, Y., Zou, Z., Chen, Y., Blissard, G.W., Kanost, M.R., and Jiang, H.
    Insect Biochem. Mol. Biol. 62,  51-63
    Full text...
  • A genome-wide analysis of antimicrobial effector genes and their transcription patterns in Manduca sexta 2015

    He, Y., Cao, X., Lia, K., Hua, Y., Chen, Y., Blissard, G., Kanost, M.R., and Jiang, H.
    Insect Biochem. Mol. Biol. 62,  23-37
    Full text...
  • Special Issue: Biological Insights from the Manduca sexta genome 2015

    Kanost, M. R., and G. W. Blissard (ed.)
    Insect Biochem. Mol. Biol. 2015/05/24 ed, vol. 62
    Full text...
  • The vacuolar protein sorting genes in insects: A comparative genome view 2015

    Li, Z., and Blissard, G.W.
    Insect Biochem. Mol. Biol. 62,  211-225
    Full text...
  • Structural features, evolutionary relationships, and transcriptional regulation of C-type lectin-domain proteins in Manduca sexta 2015

    Rao, X-J., 1, Cao, X., He, Y., Hu, Y., Zhang, X., Chen, Y., Blissard, G., Kanost, M.R., Yu, X-Q., and Jiang, H.
    Insect Biochem. Mol. Biol. 62,  75-85
    Full text...
  • Autographa californica multiple nucleopolyhedrovirus GP64 protein: Analysis of domain I and V amino acid interactions and membrane fusion activity 2015

    Yu, Q., G. W. Blissard, T. X. Liu, and Z. Li.
    Virology 488,  259-270
    Full text...
  • Analysis of chitin-binding proteins from Manduca sexta provides new insights into evolution of peritrophin A-type chitin-binding domains in insects 2015

    Tetreau, G., N. T. Dittmer, X. Cao, S. Agrawal, Y. R. Chen, S. Muthukrishnan, J. Haobo, G. W. Blissard, M. R. Kanost, and P. Wang.
    Insect Biochem. Mol. Biol. 62,  127-141
    Full text...
  • Cellular VPS4 is required for efficient entry and egress of budded virions of Autographa californica multiple nucleopolyhedrovirus 2012

    Li, Z., and Blissard, G.W.
    J. Virol. 86,  459-472
    Full text...
  • Functional analysis of the Autographa californica multiple nucleopolyhedrovirus GP64 terminal fusion loops and interactions with membranes 2012

    Dong, S., and Blissard, G.W.
    J. Virol. 86,  9617-9628
    Full text...
  • Autographa californica multiple nucleopolyhedrovirus GP64 protein: Roles of histidine residues in triggering membrane fusion and fusion pore expansion 2011

    Li, Z., and Blissard, G.W.
    J. Virol. 85,  12492-12504
    Full text...
  • Correction: BTI-Tnao38, a new cell line derived from Trichoplusia ni, is permissive for AcMNPV infection and produces high levels of recombinant proteins 2012

    Hashimoto, Y., Zhang, S., Zhang, S., Chen, Y.R., and Blissard, G.W.
    BMC Biotechnol 12:12
    Full text...
  • The Autographa californica multiple nucleopolyhedrovirus lef-5 gene is required for productive infection 2011

    Su, J., Lung, O., and Blissard, G.W.
    Virology 416,  54-64
    Full text...
  • Transcriptome responses of the host, Trichoplusia ni, to infection by the baculovirus, Autographa californica Multiple Nucleopolyhedrovirus (AcMNPV) 2014

    Chen, Y. R., Zhong, S., Fei, Z., Gao, S., Zhang, S., Li, Z., Wang, P., and Blissard, G.W.
    J. Virol. 88,,  13781-13797
    Full text...
  • Baculovirus GP64 disulfide bonds: the intermolecular disulfide bond of Autographa californica multicapsid nucleopolyhedrovirus GP64 is not essential for membrane fusion and virion budding 2010

    Li, Z., and Blissard, G.W.
    J. Virol. 84,  8584-8595
    Full text...
  • Ao38, a new cell line from eggs of the black witch moth, Ascalapha odorata (Lepidoptera: Noctuidae), is permissive for AcMNPV infection and produces high levels of recombinant proteins 2010

    Hashimoto, Y., Zhang, S., and Blissard, G.W.
    BMC Biotechnol. 10,  50
    Full text...
  • The pre-transmembrane domain of the Autographa californica multicapsid nucleopolyhedrovirus GP64 protein is critical for membrane fusion and virus infectivity 2009

    Li, Z., and Blissard, G.W.
    J. Virol. 83,  10993-11004
    Full text...
  • The Autographa californica multicapsid nucleopolyhedrovirus GP64 protein: analysis of transmembrane domain length and sequence requirements 2009

    Li, Z., and Blissard, G.W.
    J. Virol. 83,  4447-4461
    Full text...
  • Baculoviruses: Molecular Biology of Nucleopolyhedroviruses 2008

    Theilmann, D.A., and Blissard, G.W
    Desk Encyclopedia of General Virology (Mahy, B.W.J. and van Regenmortel, M. eds) Oxford, Elsevier,  434-444
    Full text...
  • Functional analysis of the transmembrane (TM) domain of the Autographa californica multicapsid nucleopolyhedrovirus GP64 protein: Substitution of heterologous TM domains 2008

    Li, Z., and Blissard, G.W.
    J. Virol. 82,  3329-3341
    Full text...
  • Display of heterologous proteins ongp64null baculovirus virions and enhanced budding mediated by a vesicular stomatitis virus G-stem construct 2008

    Zhou, J., and Blissard, G.W.
    J. Virol. 82,  1368-1377
    Full text...
  • Identification of a GP64 subdomain involved in receptor binding by budded virions of the baculovirus Autographica californica multicapsid nucleopolyhedrovirus 2008

    Zhou, J., and Blissard, G.W.
    J. Virol. 82,  4449-4460
    Full text...
  • The AcMNPV pp31 gene is not essential for productive AcMNPV replication or late gene transcription but appears to increase levels of most viral transcripts 2007

    Yamagishi, J., Burnett, E.D., Harwood, S.H., and Blissard, G.W.
    Virol. 365,  34-47
    Full text...
  • Insect cell culture and biotechnology 2007

    Granados, R.R., Li, G., and Blissard, G.W.
    Virol. Sinicia 365,  34-47
    Full text...
  • Mapping the conformational epitope of a neutralizing antibody (AcV1) directed against the AcMNPV GP64 protein 2006

    Zhou, J., and Blissard, G.W.
    Virol. 352,  427-437
    Full text...
  • On the classification and nomenclature of baculoviruses: A proposal for revision 2006

    Jehle, J.A., Blissard, G.W., Bonning, B.C., Cory, J.S., Herniou, E.A., Rohrmann, G.F., Theilmann, D.A., Thiem, S.M., and Vlak, J.M.
    Virol. 151,  1257-1266
    Full text...
  • Persistent Gene Expression in Mouse Nasal Epithelia Following Feline Immunodeficiency Virus-based Vector Gene Transfer 2005

    Sinn, P. L., Burnight, E.R., Hickey, M.A., Blissard, G.W., and McCray, P.B.
    J. Virol. 79,  12818-12827
    Full text...
  • A Cellular Drosophila melanogaster Protein with Similarity to Baculovirus F Envelope Fusion Proteins 2005

    Lung, O., and Blissard, G.W.
    J. Virol. 79,  7979-7989
    Full text...
  • Baculoviridae 2005

    Theilmann, D.A., Blissard, G.W., Bonning, B., Jehle, J., O'Reilly, D.R., Rohrmann, G.F., Thiem, S., and Vlak, J.M.
    Virus Taxonomy: Eighth Report of the International Committee on Taxonomy of Viruses (H. V. Van Regenmortel, D.H.L. Bishop, M. H. Van Regenmortal and C.M. Fauquet, eds.) Elsevier, NY 0:,  177-185
    Full text...
  • Palmitoylation of theAutographa californica multicapsid nucleopolyhedrovirus envelope glycoprotein GP64: mapping, functional studies, and lipid rafts 2003

    Zhang, S. X. X., Han, Y., and Blissard, G.W.
    J. Virol. 77,  6265-6273
    Full text...
  • Ac23, an envelope fusion protein homolog in the baculovirus Autographa californica multicapsid nucleopolyhedrovirus, is a viral pathogenicity factor 2003

    Lung, O. Y., Cruz Alvarez, M., and Blissard, G.W.
    J. Virol. 77,  328-339
    Full text...
  • Analysis of an Autographa californica nucleopolyhedrovirus lef-11 knockout: LEF-11 is essential for viral DNA replication 2002

    Lin, G., and Blissard, G.W.
    J. Virol. 76,  2770-2779
    Full text...
  • Analysis of an Autographa californica Multicapsid Nucleopolyhedrovirus lef-6-Null Virus: LEF-6 is not essential for viral replication but appears to accelerate late gene transcription 2002

    Lin, G., and Blissard, G.W.
    J. Virol. 76,  5503-5514
    Full text...
  • Pseudotyping Autographa californica Multicapsid Nucleopolyhedrovirus (AcMNPV): F proteins from group II NPVs are functionally analogous to AcMNPV GP64 2002

    Lung, O., Westenberg, M., Vlak, J.M., Zuidema, D., and Blissard, G.W.
    J. Virol. 76,  5729-5736
    Full text...
  • Stable cell lines expressing baculovirus P35: Resistance to apoptosis and nutrient stress, and increased glycoprotein secretion 2001

    Lin, G., Li, G., Granados, R.R., and Blissard, G.W.
    Vitro Cellular & Developmental Biology - Animal 37,  293-302
    Full text...
  • Expression and localization of LEF-11 in Autographa californica nucleopolyhedrovirus infected Sf9 cells 2001

    Lin, G., Slack, J.M., and Blissard, G.W.
    J. Gen. Virol. 82,  2289-2294
    Full text...
  • A GP64null baculovirus pseudotyped with Vesicular Stomatitis Virus G protein 2001

    Mangor, J. T., Monsma, S.A., Johnson, M.C., and Blissard, G.W.
    J. Virol. 75,  2544-2556
    Full text...
  • Measurement of membrane fusion activity from viral membrane fusion proteins based on a fusion-dependent promoter induction system in insect cells 2001

    Slack, J. M., and Blissard, G.W.
    J. Gen. Virol. 82,  2519-2529
    Full text...
  • A Novel Baculovirus Envelope Fusion Protein with a Proprotein Convertase Cleavage Site 2000

    IJkel, W. F. J., Westenberg, M., Goldbach, R.W., Blissard, G.W., Vlak, J.M., and Zuidema, D.
    Virol. 275,  30-41
    Full text...

Patents

    • Technology Area: Enabling Technology – Virus, Vaccines
    • Title: Baculovirus Cloning System
    • US Patent/Application(s): 5,750,383
    • Publication: J Virol 1996
    • Technology Area: Enabling Technology – Virus, Vaccines
    • Title: Baculoviruses With Enhanced Virion Production and a Method for the Production of Baculoviruses (for Retrovirus)
    • US Patent/Application(s): 13/231,324
    • Publication: J Virol 2008
    • Technology Area: Enabling Technology – Virus, Vaccines
    • Title: Baculoviruses With Enhanced Virion Production and a Method for the Production of Baculoviruses (for Baculovirus)
    • US Patent/Application(s): 12/667,956
    • Publication: J Virol 2008
    • Technology Area: Enabling Technology – Virus, Vaccines
    • Title: A GP64 Null Baculovirus Pseudotyped with Heterologous envelope proteins for gene therapy
    • US Patent/Application(s): 6,858,205
    • Publication: J Virol 2001
    • Technology Area: Enabling Technology – Virus, Vaccines
    • Title: A GP64 Null Baculovirus Pseudotyped with Heterologous envelope proteins
    • US Patent/Application(s): 6,607,912
    • Publication: J Virol 2001
    • Technology Area: Enabling Technology – Insect Cell Lines for Protein Production
    • Title: Stable Cell Resistant to Apoptosis and Nutrient Stress and Methods of Making Same (p35-Sf9 cell line)
    • US Patent/Application(s): 7,405,038
    • Publication: IVCDBA 2001
    • Additional Lead Inventor(s): Robert (Bob) Granados

Research Utilization

Research in the Blissard lab addresses fundamental and applied topics related to baculovirus infection and gene expression in insect cells. Baculoviruses are virulent pathogens of a number of insects, including many pests of agricultural importance. The baculovirus AcMNPV has been developed as a widely used expression system for secreted or membrane-bound proteins, or those requiring eukaryotic post-translational processing. Baculoviruses have been used for the development of vaccines and therapeutic proteins. They are also used to efficiently introduce DNA into mammalian cells and are being explored as potential vectors for human gene therapy. The laboratory focuses on several aspects of the baculovirus infection cycle, as well as on insect cell lines used in protein expression.

A major objective of our studies is to understand the complex nature of baculovirus entry into, and exit from insect cells. The major viral envelope protein, GP64, is critical for both attachment to host cells and fusion of viral and cellular membranes. Studies in the lab delve into the structure and function of this critical envelope protein, as well as the cellular pathways that deliver the virus to the nucleus, where viral replication occurs. The budding of baculovirus particles from infected cells is also under study. Understanding the mechanisms involved in baculovirus budding will be critically important for developing high quality virus-like particle (VLP) vaccines, which reduce or eliminate baculovirus particles from VLP preparations.

Another major objective is to understand viral gene expression, which is divided conceptually into early and late phases, mediated by host and viral RNA polymerases, respectively. The viral late RNA polymerase is critically important for biotechnological applications, as this polymerase mediates the exceptionally high levels of protein production from the baculovirus expression system, which is among the highest protein expression known in all eukaryotes. Using high throughput sequencing, we recently characterized the complete transcriptome of the baculovirus AcMNPV through the viral infection cycle in a High Five-derived cell line. In addition, we are analyzing the specific transcriptional changes and responses of the cell line to AcMNPV infection. This information will permit precise engineering of the virus and cell lines for optimized utilization in many biotechnological applications.

The laboratory also studies insect cells, focusing on the development of cell lines and most recently, on the model insect Manduca sexta. In collaboration with scientists from Kansas State University and the Baylor College of Medicine, the Manduca sexta genome was recently sequenced. Our transcriptome studies have examined gene expression in a variety of tissues over a range of developmental stages of the animal.

Recent studies are also focused on understanding how certain viruses are transmitted by insects. We are using several approaches to examine how viruses interact with and traffic through the midgut cells of vector and host insects. These studies will be important understanding how plant and animal viruses effectively utilize insects as vectors for the transmission of plant and animal diseases.

Collaborations and Consulting

In the News

Research Overview

Studies in the Blissard lab are focused on understanding the biology and pathology of viral infections and interactions in insect cells. In general, our studies can be subdivided into studies of a) baculovirus biology and pathology, b) viral envelope protein structure and function, and c) gene expression in virus infected cells.

The baculoviruses are large DNA viruses that have been used for biological control of insect pest species. In addition, baculoviruses have been developed as powerful gene expression systems for producing eukaryotic proteins in both research and industrial applications. More recently, baculoviruses have been examined as potential vectors for human gene therapy. One important thrust of our work is basic studies of the glycoproteins found in the virus envelope, and the roles of viral envelope proteins in viral attachment, entry, and exit. Another major research focus in our lab is the study of viral and host gene expression over the course of the infection. We are using next-generation sequencing technologies to examine the details of viral and host cell gene expression as infection progresses in baculovirus-infected insect cells, using both cultured cell lines and midgut tissue of host insects

Previously, we have used the baculovirus, AcMNPV, and the baculovirus GP64 envelope glycoprotein as models for our studies. We have developed AcMNPV-permissive cell lines to examine both viral envelope protein requirements, as well as cellular factors involved in entry and egress. Our recent studies suggest a model in which the promiscuous nature of baculovirus entry (which has been exploited in mammalian cell transduction systems) appears to result from recognition of, and interactions with negatively charged host membrane phospholipids. We found that this interaction appears to be mediated by specific regions of the so-called fusion loops of the major envelope glycoprotein, GP64. Our studies of GP64 have identified domains and specific amino acid positions important for virus-cell attachment, and we have also examined domains and amino acids required for the pH-triggered conformational change associated with membrane fusion during viral entry. Our studies of viral egress demonstrated that components of the cellular vacuolar protein sorting system (the cellular ESCRT pathway) are essential for baculovirus egress from infected cells. Thus, baculoviruses appear to hijack the cellular ESCRT pathway for egress from the cell.

In addition to the above studies, we are currently directing our efforts to the study of virus-insect interactions in the insect midgut. The polarized trafficking of viruses across this epithelial cell layer represents a critical virus-insect interaction that is essential for most insect-vectored viruses, but one that is not well understood in any system. Our studies focus on both identifying cellular interacting proteins and pathways that mediate polarized trafficking in the midgut, and understanding viral and cellular transcriptional responses in insect midgut cells from permissive and non-permissive hosts. We expect that our studies will have impacts on the understanding of pathogenic viruses of insects, as well as plant and animal viruses that are vectored by insects. In these studies, our goals are to generate new tools and methods for the study of the many viruses that traffic across the insect midgut epithelium.