Sindbis Virus Clone
Sindbis Virus Clone - Related Videos
07:35
Rescue and Characterization of Recombinant Virus from a New World Zika Virus Infectious Clone
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Cited by 13 •
2017
This protocol describes the recovery of infectious Zika virus from a two-plasmid infectious cDNA clone.
08:49
Propagating and Detecting an Infectious Molecular Clone of Maedi-visna Virus that Expresses Green Fluorescent Protein
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Cited by 1 •
2011
We describe a molecular clone of maedi-visna virus that expresses GFP and is fully infectious. Replication of this virus can be detected by using fluorescence microscopy and flow cytometry.
09:55
Molecular Cloning
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2023
Molecular cloning is a set of methods, which are used to insert recombinant DNA into a vector - a carrier of DNA molecules that will replicate recombinant DNA fragments in host organisms. The DNA fragment, which may be a gene, can be isolated from a prokaryotic or eukaryotic specimen. Following isolation of the fragment of interest, or insert, both the vector and insert must be cut with restriction enzymes and purified. The purified pieces are joined together though a technique called...
05:21
Culturing and Harvesting of HSPC Clones: A Method to Obtain the Singly-Sorted HSPC Clones
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2023
This video describes the protocol for isolating HSPCs from the bone marrow and sorting them using a fluorescent activated cell sorting technique, which is a method to obtain singly-sorted HSPCs. We culture these sorted cells to obtain HSPC clones.
14:23
A Restriction Enzyme Based Cloning Method to Assess the In vitro Replication Capacity of HIV-1 Subtype C Gag-MJ4 Chimeric Viruses
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Cited by 12 •
2014
HIV-1 pathogenesis is defined by both viral characteristics and host genetic factors. Here we describe a robust method that allows for reproducible measurements to assess the impact of the gag gene sequence variation on the in vitro replication capacity of the virus.
FAQs
Promoter placement upstream of the viral genome enables production of complementary RNA from the assembled construct. That RNA supplies the template for viral protein production, genome replication, and particle formation after introduction into permissive cells. Consequently, the design connects the genetic sequence selected by the researcher with the biological process used to recover virus.
Targeted mutations let investigators alter selected viral sequences while keeping the broader genetic background defined. They can then examine how those changes affect viral replication, host innate immune responses, or determinants of pathogenesis. This approach helps associate particular sequence features with observable infection-related outcomes rather than relying only on naturally occurring viral variation.
Permissive cells provide the host machinery needed after infectious RNA enters the experimental system. Viral proteins are produced, the genome is replicated, and particles are formed within those cells. Because these linked events depend on cellular support, the choice of a permissive cellular environment is central to obtaining the intended viral output from the construct.
A genetically defined clone gives researchers a controllable sequence background for comparing related viral constructs. When variants differ by selected changes, differences in replication, immune-response readouts, or pathogenicity-related behavior can be interpreted in relation to those changes. The resulting reproducibility is especially valuable when experiments support antiviral, vaccine, or virus-vector research.
The workflow begins by assembling complementary DNA that encodes the viral genome downstream of a promoter. Researchers transcribe this construct into infectious RNA and introduce the RNA into permissive cells. Cellular machinery then supports viral protein production, genome replication, and particle formation, creating material for downstream studies of alphavirus biology and infection.
These clones support several complementary questions: how viral sequences influence replication, which features contribute to pathogenesis, and how infected cells mount innate immune responses. Their controlled genetic design also supports evaluation of vaccine, antiviral, and virus-vector strategies. In immunology and infection research, the system therefore links viral genotype with cellular and disease-relevant responses.