2016년 12월 31일 토요일

Phage display

Phage display

The phage display (Phage display) is one of the methods to detect interaction between the protein or the interaction with protein and other target materials. I incorporate a gene in a bacteriophage and produce it on the surface and detect interaction for an index by the combination with the target. There is the point similar to the toe hybrid method, too, but is suitable for screening more.

A gene encoding protein and it which were displayed supports one to one in the form called the bacteriophage particle, and it is a good point that an objective gene is provided easily, and can increase. There are the in vitro virus method and ribosomal display method as the method that protein and a gene are provided to one to one likewise, but I am developed among this and others first and am the method that is used flourishingly now now.

The selection can concentrate a thing binding to the specific target from the library including a large number of genes as important application. Because I can apply this to artificial "evolution" of the protein, I liken it to natural selection (Natural selection) and say an affinity selection (affinity choice: Affinity selection) or an in vitro selection (In vitro selection).

Method

M13 bacteriophage, fd bacteriophage are used for a bacteriophage. For example, in the case of M13 bacteriophage, I connect DNA encoding objective protein or peptide with coat protein pIII or pVIII gene. I am transformed to Escherichia coli and produce the gene of this bacteriophage. Furthermore, the bacteriophage particle which displayed objective protein is released if I transmit the helper phage which has normal pIII, pVIII to form a bacteriophage particle. There is the method using Escherichia coli developing pIII, pVIII not helper phage recently.

In the affinity selection, I immobilize a target material on the surfaces of the microtiter plate and, in addition, I react it and wash the liquid including the bacteriophage more there. Then the connected bacteriophage stays, and the others are washed away. I elute a connected bacteriophage and collect it and I transmit these to Escherichia coli with helper phage and increase it. The bacteriophage which is strongly gradually connected if I repeat this is concentrated. I liken this process to "investigating gold dust from sand" and call it パンニング (panning).

Use

Though I search for specific protein and DNA and interacting protein using the DNA library which I made, I am used widely by a certain creature.

Exhaustive elucidation of the interaction between the protein can use it, but often uses fewer toe hybrid law of the false-positive for this purpose.

The method to get monoclonal antibody is developed with a phage display by the library of the antibody gene without using immunity and the hybridoma to an animal for an antigen.

In addition, protein, peptide (ペプチドアプタマー) connected using the protein which I made various kinds of modifications to and the library which, besides, encodes a random amino acid sequence to the target is used for one (man-made evolution) of improvement or that 創成 does artificially.

Allied item

This article is taken from the Japanese Wikipedia Phage display

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