What is a phagemid vector?
What is a phagemid vector?
A phagemid or phasmid is a DNA-based cloning vector, which has both bacteriophage and plasmid properties. These vectors carry, in addition to the origin of plasmid replication, an origin of replication derived from bacteriophage.
Why is pBluescript a good vector?
Gene Cloning The versatility of pBluescript vectors alleviates the need for successive subcloning. With 21 restriction sites, in two orientations, most cloned genes can be inserted directionally into the pBluescript polylinkers.
What is a phagemid a hybrid vector phage vector plasmid vector viral vector?
d) Viral vector. Explanation: Phagemid is a hybrid of M13 phage and Pbr322 plasmid. Pembl8 is an example of phagemid which was created by transferring into a pUC8 a 1300 bp fragment of M13 genome.
What is a multiple cloning site in a plasmid?
Multiple Cloning Site (MCS) Short segment of DNA which contains several restriction sites allowing for the easy insertion of DNA. In expression plasmids, the MCS is often downstream from a promoter. Insert. Gene, promoter or other DNA fragment cloned into the MCS for further study.
What are the 2 most commonly used vectors?
Two types of vectors are most commonly used: E. coli plasmid vectors and bacteriophage λ vectors. Plasmid vectors replicate along with their host cells, while λ vectors replicate as lytic viruses, killing the host cell and packaging the DNA into virions (Chapter 6).
Which type of vector is pBluescript?
The pBluescript II phagemids (plasmids with a phage origin) are cloning vectors designed to simplify commonly used cloning and sequencing procedures, including the construction of nested deletions for DNA sequencing, generation of RNA transcripts in vitro and site-specific mutagenesis and gene mapping.
What is lambda phage vector?
Enterobacteria phage λ (lambda phage, coliphage λ) is a bacterial virus, or bacteriophage, that infects the bacterial species Escherichia coli. Lambda phage consists of a virus particle including a head (also known as a capsid), tail and tail fibers. The head contains the phage’s double-stranded circular DNA genome.
What is the difference between a cloning vector and an expression vector?
In general, cloning vectors are plasmids that are used primarily to propagate DNA. An expression vector is a specialized type of cloning vector. Expression vectors are designed to allow transcription of the cloned gene and translation into protein.
What is M13 phage vector?
bacteriophage M13 – filamentous bacteriophage. single-stranded circular DNA genome (6407 bp long) – packaged inside rod-shaped protein capsid.
How do multiple cloning sites work?
A multiple cloning site (MCS), also called a polylinker, is a short segment of DNA which contains many (up to ~20) restriction sites – a standard feature of engineered plasmids. The purpose of a MCS in a plasmid is to allow a piece of DNA to be inserted into that region.
What is the role of multiple cloning sites of a vector?
A multiple cloning site (MCS, or Polylinker region) is a DNA region within a Plasmid that contains multiple unique Restriction enzyme cut sites. This enables the plasmid to act as a vector to insert DNA into another cell, for example to create a transgenic organism or for use in gene therapy.
What can a pBluescript 2 phagemid be used for?
INTRODUCTION The pBluescript II phagemids (plasmids with a phage origin) are cloning vectors designed to simplify commonly used cloning and sequencing procedures, including the construction of nested deletions for DNA sequencing, generation of RNA transcripts in vitro and site-specific mutagenesis and gene mapping.
Why is pBluescript used as a gene mapping vector?
We believe that this difficulty was also partly due to the fact that the pMG-725/EGFP-NPL plasmid has a pBluscript backbone, i.e. is a high copy plasmid that cannot accommodate a big exogenous fragment due to rearrangement during plasmid amplification in the bacteria (Alting- Mees and Short, 1989; Nakano et al., 1995).
How are phagemids used as a cloning vector?
The phagemids are cloning vectors to simplify commonly used cloning and sequencing procedures, including nested deletion construction for DNA sequencing, generation of RNA transcripts in vitro, and site-specific mutagenesis and gene mapping. The phagemids have an extensive polylinker with 21 unique restriction enzyme recognition sites.
How is a phagemid different from a plasmid?
PHAGEMID VECTORS A phagemid or phasmid is a type of cloning vector developed as a hybrid of the filamentous phage M13 and plasmids to produce a vector that can grow as a plasmid, and also be packaged as single stranded DNA in viral particles.
