Topic
Polyhedrin
About: Polyhedrin is a research topic. Over the lifetime, 1079 publications have been published within this topic receiving 40118 citations. The topic is also known as: IPR001746 & Polyhedrin.
Papers published on a yearly basis
Papers
Book•
1 Jan 1992
TL;DR: This laboratory manual simplifies selection of the most appropriate baculovirus vector design for a given problem by describing each step of the implementation process - from vector construction to large-scale protein production.
Abstract: Acknowledgments How To Use This Manual PART I: AN OVERVIEW OF BACULOVIRUSES 1: Virus Structure and the Infection Process 2: Gene Organization, Regulation, and Function 3: Virus-Host Interactions 4: Summary of Baculovirus Features Relevant to Expression Factors PART II: CHOOSING A TRANSFER PLASMID AND PARENT VIRUS 5: Choice of Virus and Host Species 6: Choice of Transfer Plasmid 7: Available Transfer Plasmids 8: Choosing a Parent Virus for Use in Vector Construction 9: Optimizing Expression: Tailoring the Heterologous Gene to the Transfer Plasmid and the Baculovirus Expression System PART III: METHODS FOR VECTOR CONSTRUCTION AND GENE EXPRESSION 10: Safety Considerations 11: Insect Cell Culture 12: Virus Methods 13: Contransfection and Recombinant Virus Identification 14: Characterizing Recombinant Gene Expression 15: Post-Translational Modification PART IV: METHODS FOR SCALE-UP OF PROTEIN PRODUCTION AND USE OF INSECT LARVAE 16: David L. Clemm: Scale-Up of Protein Production in a Stirred Bioreactor 17: Cheryl Isaac Murphy: Scale-Up of Protein Production in an Airlift Fermenter 18: Insect Rearing and Infection Appendix 1: Nucleotide Sequence of AcMNPV Polyhedrin Region Appendix 2: Nucleotide Sequence of AcMNPV p10 Region Appendix 3: List of Genes Expressed in the Baculovirus Expression System Appendix 4: Suppliers Appendix 5: Stock Solutions Appendix 6: Excel Spreadsheet for TCID[5[0 Calculation References Index
2,461 citations
1 May 1987
1,990 citations
TL;DR: The current status and potential use of baculovirus vectors for the expression of foreign genes in insect cells are described and are contributing to understanding the molecular biology of gene and protein function and regulation in both vertebrate and insect systems.
Abstract: We describe the current status and potential use of baculovirus vectors for the expression of foreign genes in insect cells. Trends in the development of transfer vectors for the expression of foreign genes under the control of the strong polyhedrin promoter of Autographa californica nuclear polyhedrosis virus and strategies for maximizing levels of expression are discussed. Baculovirus vectors have achieved widespread acceptance for their ability to express proteins of agricultural and medical importance. A baculovirus vector was used to express the first recombinant HIV envelope proteins to receive F.D.A. approval for clinical evaluation as a candidate vaccine for AIDS. These insect DNA virus vectors are contributing to understanding the molecular biology of gene and protein function and regulation in both vertebrate and insect systems.
1,191 citations
TL;DR: Results demonstrate that AcNPV should be suitable for use as a eucaryotic expression vector for the production of products from cloned genes.
Abstract: Autographa californica nuclear polyhedrosis virus (AcNPV) was used as an expression vector for human beta interferon. By using specially constructed plasmids, the protein-coding sequences for interferon were linked to the AcNPV promoter for the gene encoding for polyhedrin, the major occlusion protein. The interferon gene was inserted at various locations relative to the AcNPV polyhedrin transcriptional and translational signals, and the interferon-polyhedrin hybrid genes were transferred to infectious AcNPV expression vectors. Biologically active interferon was produced, and greater than 95% was secreted from infected insect cells. A maximum of ca. 5 X 10(6) U of interferon activity was produced by 10(6) infected cells. These results demonstrate that AcNPV should be suitable for use as a eucaryotic expression vector for the production of products from cloned genes.
1,169 citations
TL;DR: The requirements for high level expression of three foreign proteins using the polyhedrin gene promoter of Autographa californica nuclear polyhedrosis virus (AcNPV, Baculoviridae) have been investigated and it has been estimated that LCMV N protein represented approximately 50% of the total cellular protein, an observation consistent with the presence of numerous inclusion bodies in the cytoplasm of infected cells.
Abstract: Summary
The requirements for high level expression of three foreign proteins using the polyhedrin gene promoter of Autographa californica nuclear polyhedrosis virus (AcNPV, Baculoviridae) have been investigated. In Spodoptera frugiperda cells infected with the appropriate recombinant baculoviruses, the synthesis of the two S RNA coded genes of lymphocytic choriomeningitis virus (LCMV; i.e. the nucleoprotein, N, and glycoprotein precursor, GPC), or the haemagglutinin gene of influenza A virus, appears to be related to the degree of integrity of the 5′ upstream sequence of the polyhedrin gene. No effect on the level of N protein expression was detected when all the polyhedrin gene coding sequences or some of the immediate 3′ downstream sequences were deleted. Using the most efficient expression viruses derived from a new transfer vector, pAcYM1, it has been estimated that LCMV N protein represented approximately 50% of the total cellular protein, an observation consistent with the presence of numerous inclusion bodies in the cytoplasm of infected cells. For recombinant viruses derived from the pAcYM1 transfer vector containing the LCMV GPC gene, the level of synthesis of the arenavirus glycoprotein was equivalent to approximately 20% of the cellular protein. Thin sections of cells infected with the GPC recombinant revealed a highly vacuolated cytoplasm.
732 citations
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