Phage Vector Pdf

Lambda phage consists of a virus particle including a head also known as a capsid tail and tail fibers.

Phage vector pdf. 40204024 april 1996 chemistry mapping proteinprotein interactions by affinity directed mass spectrometry. It has the following advantages. The vector therefore contains features that allow for the convenient insertion or removal of a dna fragment to. Stoker ng grant ka dockrell hm howard cr jouy nf mcadam kp.

The cosmid vector can carry up to 45 kb whereas plasmid and l phage vectors are limited to 25 kb. High level expression of genes cloned in phage lambda gt11. The plasmid vector is cleaved to generate two vector arms to which up to 100 kb of foreign dna is ligated and packaged into a pi protein coat in vitro. Clontech mountain view ca was amplified by pcr attaching noti and bamh1 restriction sites to 5 and 3 ends respectively.

A cloning vector is a small piece of dna that can be stably maintained in an organism and into which a foreign dna fragment can be inserted for cloning purposes. Single stranded forms of the phage. Cdna encoding a variant of the red fluorescent protein adapted from discosoma sp. The cloning vector may be dna taken from a virus the cell of a higher organism or it may be the plasmid of a bacterium.

The head contains the phages double stranded circular dna genome. Cloning into m13mp vectors. The phage vector was modified for dual transgenesis as follows. This recombinant pi phage is then allowed to adsorb to a suitable host following which the recombinant pi dna is injected into the cell circularized and are amplified fig.

Lowman in encyclopedia of biological chemistry second edition 2013. Sternberg n tiemeier d enquist l. Development of m13 into a cloning vector. In vitro packaging of a lambda dam vector containing ecori dna fragments of escherichia coli and phage p1.

Phagemid vectors consist of a typical plasmid construct into which a phage origin of replication ori including the phage packaging region of dna from the phage genome and a gene for the displayed fusion protein have been inserted figure 1in this case the phagemid dna can be replicated. Cosmid vector the cosmid vector is a combination of the plasmid vector and the cos site which allows the target dna to be inserted into the l head. This simplifies the intermediate amplification steps during. Additionally the vector does not carry a plasmid replicon or antibiotic resistance so it is necessary to propagate the vector as phage rather than a plasmid ie titer for plaques not colonies.

This amplicon was cloned into the phage backbone in the. The phage particle recognizes and binds to its host e.