Hbv Genome Plasmids
Hbv Genome Plasmids - Related Videos
08:16
Plasmid Purification
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2023
Plasmid purification is a technique used to isolate and purify plasmid DNA from genomic DNA, proteins, ribosomes, and the bacterial cell wall. A plasmid is a small, circular, double-stranded DNA that is used as a carrier of specific DNA molecules. When introduced into a host organism via transformation, a plasmid will be replicated, creating numerous copies of the DNA fragment under study. In this video, a step-by-step generalized procedure is described for how to perform plasmid purification.
08:31
Genome Editing
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2023
A well-established technique for modifying specific sequences in the genome is gene targeting by homologous recombination, but this method can be laborious and only works in certain organisms. Recent advances have led to the development of “genome editing”, which works by inducing double-strand breaks in DNA using engineered nuclease enzymes guided to target genomic sites by either proteins or RNAs that recognize specific sequences. When a cell attempts to repair this damage, mutations can be...
08:31
Isolation and Genome Analysis of Single Virions using 'Single Virus Genomics'
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Cited by 2 •
2013
Single Virus Genomics (SVG) is a method to isolate and amplify the genomes of single virons. Viral suspensions of a mixed assemblage are sorted using flow cytometry onto a microscope slide with discrete wells containing agarose, thereby capturing the virion and reducing genome shearing during downstream processing. Whole genome amplification is achieved using multiple displacement amplification (MDA) resulting in genomic material that is suitable for sequencing.
02:44
Generation of Recombinant Adeno-Associated Virus Through Plasmid Transfection
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2026
Source: Ding, J., et al. Preparation of rAAV9 to Overexpress or Knockdown Genes in Mouse Hearts. J. Vis. Exp. (2016)This video demonstrates the production of recombinant adeno-associated virus particles through plasmid transfection in mammalian cells. It highlights the coordinated expression of Rep, Cap, and helper proteins that drive viral genome replication and capsid assembly within the nucleus.
02:22
Production of Pseudovirus Using Plasmid Transfection in Human Cells
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2026
Source: Chang, X., et al. Preparation of Pseudo-Typed H5 Avian Influenza Viruses with Calcium Phosphate Transfection Method and Measurement of Antibody Neutralizing Activity. J. Vis. Exp. (2021)This video demonstrates the production of influenza pseudoviruses in human cells using calcium phosphate-mediated plasmid transfection, enabling safe generation of single-cycle viral particles for downstream applications such as antibody neutralization assays under biosafety level 2 conditions.
FAQs
Following transfection, host transcription machinery produces pregenomic RNA, which serves as a central intermediate for viral protein synthesis. In suitable cellular systems, this RNA also supports nucleocapsid formation and reverse transcription of the viral genome. Measuring outcomes linked to these steps helps investigators distinguish effects on transcription, protein production, particle assembly, or genome replication.
Defined genomic portions allow investigators to focus on selected viral functions without necessarily modeling every activity supported by a complete construct. This design can help examine antigen expression, specific viral mutations, or particular virus–cell interactions. Comparing constructs with different genomic content reveals which sequences contribute to an observed replication, immune, or cellular response.
The cellular system and the portion of the viral genome carried by the construct strongly influence the resulting readouts. Transfection must occur in cells capable of supporting the relevant activity, because nucleocapsid formation and reverse transcription occur only in suitable systems. Consequently, antigen expression, replication-associated signals, and host responses may not all be observed under identical conditions.
A typical workflow begins by selecting a construct containing the complete genome or a defined region, introducing it into susceptible cells by transfection, and examining the resulting viral or cellular readouts. Investigators can then assess viral RNA and protein expression, and, when the system supports them, nucleocapsid formation and genome reverse transcription. This creates a controlled framework for comparing experimental conditions.
Researchers can introduce the plasmid into a controlled cell system, expose the resulting model to an antiviral compound, and compare viral or cellular readouts with untreated conditions. Depending on the construct and cells, measurements may include viral RNA, protein expression, nucleocapsid formation, or genome reverse transcription. These comparisons help identify effects on defined stages of the modeled viral process.
Plasmid-based systems permit controlled analysis of selected viral mutations and their effects on replication-related activities, antigen expression, or interactions with host cells. They also provide a reproducible setting for examining immune responses associated with viral activity. In immunology and infection research, these observations can help investigate mechanisms that may contribute to persistent hepatitis B infection.