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A Time and Cost-Effective Pipeline for Expression Screening and Protein Production in Insect Cells Based on the HR-Bac Toolbox to Generate Recombinant Baculoviruses
The baculovirus expression vector system (BEVS) is recognized as a powerful platform for producing challenging proteins and multiprotein complexes...
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Adaptive Laboratory Evolution to Improve Recombinant Protein Production Using Insect Cells
Adaptive laboratory evolution (ALE) is a powerful tool for enhancing the fitness of cell lines in specific applications, including recombinant...
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Expansion and Storage of Insect Cells and Virus Stocks
Healthy insect cell cultures are critical for any method described in this book, including making productive baculovirus banks, protein or AAV...
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Recombinant AAV Production
The insect cell-baculovirus expression vector (IC-BEV) platform has enabled small research-scale and large commercial-scale production of recombinant...
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Functional Baculovirus Particle Quantification via Plaque Assay
Plaque assay method enables the quantification of infectious baculovirus when defined as plaque forming units (PFU). It allows to determine the...
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TCID50 Assay: A Simple Method to Determine Baculovirus Titer
There are many methods that can be used to determine the infectious titer of your baculovirus stock. The TCID50 method is a simple end-point dilution...
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RNAi-Mediated Silencing in the Insect Cell-Baculovirus Expression System
RNA interference (RNAi) serves as an indispensable tool for gene function studies and has been substantiated through extensive research for its...
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In Situ Protein Microarray for Identifying the Geminivirus–Arabidopsis Interactome
The protein–protein interactions (PPI) by protein array technology complement other PPI assay technologies such as AP-MS and Y2H. The in situ protein...
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Experimental Transmission of Plant Viruses by Aphids or Whiteflies
Numerous species of plant viruses are naturally transmitted by insect vectors, mainly homopterans like aphids and whiteflies. Depending on the vector...
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Quantification of Virion-Sense and Complementary-Sense DNA Strands of Circular Single-Stranded DNA Viruses
Circular ssDNA viruses are ubiquitous and can be found in both prokaryotes and eukaryotes. To understand the interaction of ssDNA viruses with their...
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Begomovirus Inoculation in Arabidopsis and Cassava
The use of infectious clones to inoculate plant viruses allows for controlled studies that lead to a better understanding of plant–virus...
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Replication Assay of Begomovirus in Arabidopsis Protoplasts
Protoplasts are isolated plant cells from which the cell walls have been removed by treatment with fungal cellulase and macerozyme enzymes, which...
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SARS-CoV-2 Detection and Genome Sequencing in Urban Wastewaters
Due to the excretion of SARS-CoV-2 in faeces, the use of wastewater-based epidemiology (WBE) is a useful tool for virus surveillance in large...
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From Whole Genome Shotgun Sequencing to Viral Community Profiling: The ViromeScan Tool
ViromeScan is an innovative metagenomic analysis tool that allows characterizing the taxonomy of viral communities from raw data of metagenomics...
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Targeted Whole Genome Sequencing of the Capripoxvirus Genome from Clinical Tissue Samples and Lyophilized Vaccine Batches
Diseases caused by Capripoxviruses (CaPVs) are of great economic importance in sheep, goats, and cattle. Since CaPV strains are serologically...
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Freshwater Viral Metagenome Analyses Targeting dsDNA Viruses
Viral metagenomics is one of the most widely used approaches to study viral population genomics. With the recent development of bioinformatic tools,...
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Observation of Bacteriophage Ultrastructure by Cryo-Electron Microscopy
Transmission electron microscopy (TEM) is an ideal method to observe and determine the structure of bacteriophages. From early studies by negative...
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Reverse Genetics of Zika Virus Using a Bacterial Artificial Chromosome
Zika virus (ZIKV) is a mosquito-borne member of the Flaviviridae family that has become a global threat to human health. Although ZIKV has been known...
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Guidelines to Compose an Ideal Bacteriophage Cocktail
Properly designed bacteriophage therapeutics are the cornerstone for a successful outcome of bacteriophage therapy. Here we present an overview of...
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Bacteriophage Production in Compliance with Regulatory Requirements
In this chapter, we discuss production requirements for therapeutic bacteriophage preparations. We review the current regulatory expectancies and...