What are Phasmid vectors?

What are Phasmid vectors?

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.

What is the difference between phagemid and plasmid?

is that plasmid is (cytology) a loop of double-stranded dna that is separate from and replicates independently of the chromosomes, most commonly found in bacteria]], but also in archaeans and [[eukaryote|eukaryotic cells, and used in genetic engineering as a vector for gene transfer while phagemid is a phage whose …

What is Hyperphage?

Hyperphage is a helper phage, which does not provide any wild type p3 for particle assembly and therefore, all p3 originates from the phagemid leading to forced oligovalent display of the protein of interest [15].

What does helper phage do?

Helper phages (e.g. M13K07) provide all the necessary gene products for particle formation when using phagemid vectors. They are mutated wild‐type phage containing the whole genome, with a defective origin of replication or packaging signal, and hence, are inefficient in self‐packaging.

What is a Fosmid library?

Fosmids are DNA vectors that use the F-plasmid origin of replication and partitioning mechanisms to allow cloning of large DNA fragments. A library that provides 20–70-fold redundant coverage of the genome can easily be prepared.

How do you propagate helper phage?

To propagate (amplify) the VCSM13 helper phage requires using the E. coli TG1, ER2738 or strainXL1-Blue MRF′, because those strain harbors the supE44 mutation, which provides a glutamine suppressor tRNA. The VCSM13 helper phage is used strictly to produce single stranded DNA from the already excised phagemid.

How do you make helper phage?

Infect 200 mg/mL XLI-Blue at an OD600 of 0.4 with 10 mL of 100-fold serial dilutions of M13KO7 helper phage in a 37 ℃ water bath for 30 min. Add to 3 mL molten top agar (50 ℃) and pour onto warm LB plates. Allow to set and incubate overnight at 37 ℃.

What is phage vector?

Phage vectors consist of an essentially complete phage genome, often M13 phage, into which is inserted DNA encoding the protein or peptide of interest (Figure 1). Typically, the remainder of the phage genome is left unchanged and provides the other gene products needed for the phage life cycle.

What is Amphid and Phasmid?

Amphids: These are the cuticular depressions present on the lips surrounding the mouth in nematodes such as Aphasmidia animals and serve as Chemoreceptors. Phasmids: These are the well developed sensory organs and they occur in some nematodes such as phasmidia animals.