Plasmid Vector
Plasmid Vector - Related Videos
08:16
Plasmid Purification
0 Views •
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.
02:44
Generation of Recombinant Adeno-Associated Virus Through Plasmid Transfection
0 Views •
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.
10:42
A Protocol for the Production of Integrase-deficient Lentiviral Vectors for CRISPR/Cas9-mediated Gene Knockout in Dividing Cells
0 Views •
Cited by 21 •
2017
We describe the production strategy of integrase-deficient lentiviral vectors (IDLVs) as vehicles for delivering CRISPR/Cas9 to cells. With an ability to mediate quick and robust gene editing in cells, IDLVs present a safer and equally effective vector platform for gene delivery compared to integrase-competent vectors.
01:28
Plasmids
0 Views •
2025
Plasmids are extrachromosomal DNA molecules found in bacteria, archaea, and some eukaryotic microbes like yeast. These small, circular DNA structures typically contain fewer than 30 genes, although some may exist linearly. Plasmids vary in their number within a cell, known as copy number. Single-copy plasmids are present in one copy per cell and multi-copy plasmids are present in multiple copies, reaching over 100 copies per cell.Plasmids usually replicate independently of the chromosomal DNA...
07:11
Homemade Site Directed Mutagenesis of Whole Plasmids
0 Views •
Cited by 91 •
2009
Site directed mutagenesis of whole plasmids is a simple way to create slightly different variations of an original plasmid. Here we demonstrate an easy and cost effective way to introduce base substitutions into a plasmid using standard reagents.
FAQs
The origin of replication is the sequence that enables a plasmid vector to duplicate separately from the host chromosome. Its function depends on compatibility with the host cell, so a plasmid may replicate effectively in one host but not another. This compatibility determines whether the introduced genetic material can be maintained during gene cloning or expression studies.
Selectable markers help researchers identify cells associated with the desired plasmid, while multiple cloning sites provide defined locations for inserting target DNA sequences. These features address different stages of recombinant DNA construction: selection supports identification, and the cloning site supports sequence insertion. Together, they make plasmid-based manipulation more organized and experimentally useful.
A promoter directs transcription of an inserted gene, making it essential when the goal is gene expression rather than only DNA cloning. Its activity must correspond to the host system, because expression vectors may be designed for bacterial or eukaryotic cells. Promoter choice therefore affects whether the target sequence is transcribed in the intended biological context.
A typical workflow begins by selecting a plasmid with features suited to the intended host and purpose. Researchers then insert the target sequence at an appropriate multiple cloning site, use the selectable marker to identify relevant cells, and rely on the origin of replication to maintain the construct. Expression-focused work also requires a promoter suited to the host.
For gene cloning, the priority is constructing and maintaining a plasmid containing the target sequence, with selection and replication supporting recovery of the recombinant DNA. Protein production additionally requires expression control, including a promoter that directs transcription in the chosen host. This distinction lets researchers match vector design to whether they need DNA analysis or gene-derived protein.
Plasmid vectors provide a flexible platform for recombinant DNA construction, protein production, and gene function studies. In biology, they help researchers manipulate and analyze selected genes within compatible cellular systems. In biotechnology, the same design principles support the production or study of gene products, while selectable markers, cloning sites, replication origins, and promoters provide the necessary control points.